Publications by authors named "Monica Monici"

29 Publications

  • Page 1 of 1

Effect of NIR Laser Therapy by MLS-MiS Source on Fibroblast Activation by Inflammatory Cytokines in Relation to Wound Healing.

Biomedicines 2021 Mar 16;9(3). Epub 2021 Mar 16.

ASA Campus Joint Laboratory, ASA Research Division & Department of Experimental and Clinical Biomedical Sciences "Mario Serio", University of Florence, I-50139 Florence, Italy.

The fine control of inflammation following injury avoids fibrotic scars or impaired wounds. Due to side effects by anti-inflammatory drugs, the research is continuously active to define alternative therapies. Among them, physical countermeasures such as photobiomodulation therapy (PBMT) are considered effective and safe. To study the cellular and molecular events associated with the anti-inflammatory activity of PBMT by a dual-wavelength NIR laser source, human dermal fibroblasts were exposed to a mix of inflammatory cytokines (IL-1β and TNF-α) followed by laser treatment once a day for three days. Inducible inflammatory key enzymatic pathways, as iNOS and COX-2/mPGES-1/PGE2, were upregulated by the cytokine mix while PBMT reverted their levels and activities. The same behavior was observed with the proangiogenic factor vascular endothelial growth factor (VEGF), involved in neovascularization of granulation tissue. From a molecular point of view, PBMT retained NF-kB cytoplasmatic localization. According to a change in cell morphology, differences in expression and distribution of fundamental cytoskeletal proteins were observed following treatments. Tubulin, F-actin, and α-SMA changed their organization upon cytokine stimulation, while PBMT reestablished the basal localization. Cytoskeletal rearrangements occurring after inflammatory stimuli were correlated with reorganization of membrane α5β1 and fibronectin network as well as with their upregulation, while PBMT induced significant downregulation. Similar changes were observed for collagen I and the gelatinolytic enzyme MMP-1. In conclusion, the present study demonstrates that the proposed NIR laser therapy is effective in controlling fibroblast activation induced by IL-1β and TNF-α, likely responsible for a deleterious effect of persistent inflammation.
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http://dx.doi.org/10.3390/biomedicines9030307DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8002295PMC
March 2021

Evaluation of Adjunctive Photobiomodulation (PBMT) for COVID-19 Pneumonia via Clinical Status and Pulmonary Severity Indices in a Preliminary Trial.

J Inflamm Res 2021 19;14:965-979. Epub 2021 Mar 19.

Department of Orthopedics, Lowell General Hospital, Lowell, MA, 01863, USA.

Purpose: Evidence-based and effective treatments for COVID-19 are limited, and a new wave of infections and deaths calls for novel, easily implemented treatment strategies. Photobiomodulation therapy (PBMT) is a well-known adjunctive treatment for pain management, wound healing, lymphedema, and cellulitis. PBMT uses light to start a cascade of photochemical reactions that lead to local and systemic anti-inflammatory effects at multiple levels and that stimulate healing. Numerous empirical studies of PBMT for patients with pulmonary disease such as pneumonia, COPD and asthma suggest that PBMT is a safe and effective adjunctive treatment. Recent systematic reviews suggest that PBMT may be applied to target lung tissue in COVID-19 patients. In this preliminary study, we evaluated the effect of adjunctive PBMT on COVID-19 pneumonia and patient clinical status.

Patients And Methods: We present a small-scale clinical trial with 10 patients randomized to standard medical care or standard medical care plus adjunctive PBMT. The PBMT group received four daily sessions of near-infrared light treatment targeting the lung tissue via a Multiwave Locked System (MLS) laser. Patient outcomes were measured via blood work, chest x-rays, pulse oximetry and validated scoring tools for pneumonia.

Results: PBMT patients showed improvement on pulmonary indices such as SMART-COP, BCRSS, RALE, and CAP (Community-Acquired Pneumonia questionnaire). PBMT-treated patients showed rapid recovery, did not require ICU admission or mechanical ventilation, and reported no long-term sequelae at 5 months after treatment. In the control group, 60% of patients were admitted to the ICU for mechanical ventilation. The control group had an overall mortality of 40%. At a 5-month follow-up, 40% of the control group experienced long-term sequelae.

Conclusion: PBMT is a safe and effective potential treatment for COVID-19 pneumonia and improves clinical status in COVID-19 pneumonia.
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http://dx.doi.org/10.2147/JIR.S301625DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7989376PMC
March 2021

A 57-Year-Old African American Man with Severe COVID-19 Pneumonia Who Responded to Supportive Photobiomodulation Therapy (PBMT): First Use of PBMT in COVID-19.

Am J Case Rep 2020 Aug 15;21:e926779. Epub 2020 Aug 15.

Department of Biological Sciences, University of Lethbridge, Lethbridge, AB, Canada.

BACKGROUND Coronavirus disease 2019 (COVID-19) is associated with lung inflammation and cytokine storm. Photobiomodulation therapy (PBMT) is a safe, non-invasive therapy with significant anti-inflammatory effects. Adjunct PBMT has been employed in treating patients with lung conditions. Human studies and experimental models of respiratory disease suggest PBMT reduces inflammation and promotes lung healing. This is the first time supportive PBMT was used in a severe case of COVID-19 pneumonia. CASE REPORT A 57-year-old African American man with severe COVID-19 received 4 once-daily PBMT sessions by a laser scanner with pulsed 808 nm and super-pulsed 905 nm modes for 28 min. The patient was evaluated before and after treatment via radiological assessment of lung edema (RALE) by CXR, pulmonary severity indices, blood tests, oxygen requirements, and patient questionnaires. Oxygen saturation (SpO₂) increased from 93-94% to 97-100%, while the oxygen requirement decreased from 2-4 L/min to 1 L/min. The RALE score improved from 8 to 5. The Pneumonia Severity Index improved from Class V (142) to Class II (67). Additional pulmonary indices (Brescia-COVID and SMART-COP) both decreased from 4 to 0. CRP normalized from 15.1 to 1.23. The patient reported substantial improvement in the Community-Acquired Pneumonia assessment tool. CONCLUSIONS This report has presented supportive PBMT in a patient with severe COVID-19 pneumonia. Respiratory indices, radiological findings, oxygen requirements, and patient outcomes improved over several days and without need for a ventilator. Future controlled clinical trials are required to evaluate the effects of PBMT on clinical outcomes in patients with COVID-19 pneumonia.
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http://dx.doi.org/10.12659/AJCR.926779DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7449510PMC
August 2020

Magnetic fields modulate metabolism and gut microbiome in correlation with Pgc-1α expression: Follow-up to an in vitro magnetic mitohormetic study.

FASEB J 2020 08 6;34(8):11143-11167. Epub 2020 Jul 6.

Department of Surgery, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore.

Exercise modulates metabolism and the gut microbiome. Brief exposure to low mT-range pulsing electromagnetic fields (PEMFs) was previously shown to accentuate in vitro myogenesis and mitochondriogenesis by activating a calcium-mitochondrial axis upstream of PGC-1α transcriptional upregulation, recapitulating a genetic response implicated in exercise-induced metabolic adaptations. We compared the effects of analogous PEMF exposure (1.5 mT, 10 min/week), with and without exercise, on systemic metabolism and gut microbiome in four groups of mice: (a) no intervention; (b) PEMF treatment; (c) exercise; (d) exercise and PEMF treatment. The combination of PEMFs and exercise for 6 weeks enhanced running performance and upregulated muscular and adipose Pgc-1α transcript levels, whereas exercise alone was incapable of elevating Pgc-1α levels. The gut microbiome Firmicutes/Bacteroidetes ratio decreased with exercise and PEMF exposure, alone or in combination, which has been associated in published studies with an increase in lean body mass. After 2 months, brief PEMF treatment alone increased Pgc-1α and mitohormetic gene expression and after >4 months PEMF treatment alone enhanced oxidative muscle expression, fatty acid oxidation, and reduced insulin levels. Hence, short-term PEMF treatment was sufficient to instigate PGC-1α-associated transcriptional cascades governing systemic mitohormetic adaptations, whereas longer-term PEMF treatment was capable of inducing related metabolic adaptations independently of exercise.
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http://dx.doi.org/10.1096/fj.201903005RRDOI Listing
August 2020

Effect of Unloading Condition on the Healing Process and Effectiveness of Platelet Rich Plasma as a Countermeasure: Study on In Vivo and In Vitro Wound Healing Models.

Int J Mol Sci 2020 Jan 9;21(2). Epub 2020 Jan 9.

ASA campus Joint Laboratory, ASA Res. Div., Department of Experimental and Clinical Biomedical Sciences "Mario Serio", University of Florence, 50139 Florence, Italy.

Wound healing is a very complex process that allows organisms to survive injuries. It is strictly regulated by a number of biochemical and physical factors, mechanical forces included. Studying wound healing in space is interesting for two main reasons: (i) defining tools, procedures, and protocols to manage serious wounds and burns eventually occurring in future long-lasting space exploration missions, without the possibility of timely medical evacuation to Earth; (ii) understanding the role of gravity and mechanical factors in the healing process and scarring, thus contributing to unravelling the mechanisms underlying the switching between perfect regeneration and imperfect repair with scarring. In the study presented here, a new in vivo sutured wound healing model in the leech () has been used to evaluate the effect of unloading conditions on the healing process and the effectiveness of platelet rich plasma (PRP) as a countermeasure. The results reveal that microgravity caused a healing delay and structural alterations in the repair tissue, which were prevented by PRP treatment. Moreover, investigating the effects of microgravity and PRP on an in vitro wound healing model, it was found that PRP is able to counteract the microgravity-induced impairment in fibroblast migration to the wound site. This could be one of the mechanisms underlying the effectiveness of PRP in preventing healing impairment in unloading conditions.
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http://dx.doi.org/10.3390/ijms21020407DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7013931PMC
January 2020

Effect of NIR laser therapy by MLS-MiS source against neuropathic pain in rats: in vivo and ex vivo analysis.

Sci Rep 2019 06 26;9(1):9297. Epub 2019 Jun 26.

Department of Neuroscience, Psychology, Drug Research and Child Health - NEUROFARBA - Pharmacology and Toxicology Section, University of Florence, Florence, Italy.

Neuropathic pain is characterized by an uncertain etiology and by a poor response to common therapies. The ineffectiveness and the frequent side effects of the drugs used to counteract neuropathic pain call for the discovery of new therapeutic strategies. Laser therapy proved to be effective for reducing pain sensitivity thus improving the quality of life. However, its application parameters and efficacy in chronic pain must be further analyzed. We investigated the pain relieving and protective effect of Photobiomodulation Therapy in a rat model of compressive mononeuropathy induced by Chronic Constriction Injury of the sciatic nerve (CCI). Laser (MLS-MiS) applications started 7 days after surgery and were performed ten times over a three week period showing a reduction in mechanical hypersensitivity and spontaneous pain that started from the first laser treatment until the end of the experiment. The ex vivo analysis highlighted the protective role of laser through the myelin sheath recovery in the sciatic nerve, inhibition of iNOS expression and enhancement of EAAT-2 levels in the spinal cord. In conclusion, this study supports laser treatment as a future therapeutic strategy in patients suffering from neuropathic pain induced by trauma.
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http://dx.doi.org/10.1038/s41598-019-45469-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6594937PMC
June 2019

Glyoxylate cycle activity in Pinus pinea seeds during germination in altered gravity conditions.

Plant Physiol Biochem 2019 Jun 3;139:389-394. Epub 2019 Apr 3.

Department of Experimental and Clinical Biomedical Sciences "Mario Serio", University of Florence, Viale Pieraccini 6, I-50139, Florence, Italy.

This work inserts in the research field regarding the effects of altered gravity conditions on biological plant processes. Pinus pinea seeds germination was studied in simulated microgravity (2x10g) and hypergravity (20g) conditions. The effects of simulated gravity were evaluated monitoring the levels of the key enzymes, involved in the main metabolic pathway during germination process of lipid-rich seeds (oilseeds): isocitrate lyase and malate synthase for glyoxylate cycle, 3-hydroxyacyl-CoA dehydrogenase for beta-oxidation, isocitrate dehydrogenase for Krebs cycle, pyruvate kinase for glycolysis and glucose 6 phosphate dehydrogenase for pentose phosphate shunt. The simulated micro and hypergravity conditions were obtained by a Random Position Machine and a Hyperfuge, respectively. Results show that the levels of some tested enzymes, at different lag times of the germination process, have the same trend of controls (g = 1), but with significant differences from quantitative point of view. They are higher in microgravity conditions and lower in hypergravity ones, suggesting that, from a biochemical point of view, the germination process results accelerated in microgravity conditions and delayed in hypergravity ones. These biochemical results show a good correlation with morphological ones, obtained with the measurement of the length of the seeds sprouting radicle. These results give promising indications regarding the possibility to grow plant with lipid-rich seeds in spatial environment, to obtain food sources for astronauts during long term space missions and to reconstitute new atmosphere.
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http://dx.doi.org/10.1016/j.plaphy.2019.03.042DOI Listing
June 2019

Continuous Exposure to Simulated Hypergravity-Induced Changes in Proliferation, Morphology, and Gene Expression of Human Tendon Cells.

Stem Cells Dev 2018 06 17;27(12):858-869. Epub 2018 Apr 17.

1 3B's Research Group-Biomaterials, Biodegradables and Biomimetics, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, University of Minho , Guimarães, Portugal .

Gravity influences physical and biological processes, especially during development and homeostasis of several tissues in the human body. Studies under altered gravity have been receiving great attention toward a better understanding of microgravity-, hypogravity (<1 g)-, or hypergravity (>1 g)-induced alterations. In this work, the influence of simulated hypergravity over human tendon-derived cells (hTDCs) was studied at 5, 10, 15, and 20 g for 4 or 16 h, using a large diameter centrifuge. Main results showed that 16 h of simulated hypergravity limited cell proliferation. Cell area was higher in hTDCs cultured at 5, 10, and 15 g for 16 h, in comparison to 1 g control. Actin filaments were more pronounced in hTDCs cultured at 5 and 10 g for 16 h. Focal adhesion kinase (FAK) was mainly expressed in focal adhesion sites upon hypergravity stimulation, in comparison to perinuclear localization in control cells after 16 h; and FAK number/cell increased with increasing g-levels. A tendency toward an upregulation of tenogenic markers was observed; scleraxis (SCX), tenascin C (TNC), collagen type III (COL3A1), and decorin (DCN) were significantly upregulated in hTDCs cultured at 15 g and COL3A1 and DCN were significantly upregulated in hTDCs cultured at 20 g. Overall, simulated hypergravity affected the behavior of hTDCs, with more pronounced effects in the long-term period (16 h) of stimulation.
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http://dx.doi.org/10.1089/scd.2017.0206DOI Listing
June 2018

Preparation of A Spaceflight: Apoptosis Search in Sutured Wound Healing Models.

Int J Mol Sci 2017 Dec 3;18(12). Epub 2017 Dec 3.

Department of Biomedicine, Aarhus University, 8000 Aarhus C, Denmark.

To prepare the ESA (European Space Agency) spaceflight project "Wound healing and Sutures in Unloading Conditions", we studied mechanisms of apoptosis in wound healing models based on ex vivo skin tissue cultures, kept for 10 days alive in serum-free DMEM/F12 medium supplemented with bovine serum albumin, hydrocortisone, insulin, ascorbic acid and antibiotics at 32 °C. The overall goal is to test: (i) the viability of tissue specimens; (ii) the gene expression of activators and inhibitors of apoptosis and extracellular matrix components in wound and suture models; and (iii) to design analytical protocols for future tissue specimens after post-spaceflight download. Hematoxylin-Eosin and Elastica-van-Gieson staining showed a normal skin histology with no signs of necrosis in controls and showed a normal wound suture. TdT-mediated dUTP-biotin nick end labeling for detecting DNA fragmentation revealed no significant apoptosis. No activation of caspase-3 protein was detectable. , , , , , , , , , and mRNAs were not altered in epidermis and dermis samples with and without a wound compared to 0 day samples (specimens investigated directly post-surgery). , , and mRNAs were downregulated in epidermis/dermis samples with and/or without a wound compared to 0 day samples. , were upregulated in 10 day wound samples compared to 0 day samples in epidermis/dermis. mRNAs were elevated in 10 day wound and no wound samples compared to 0 day samples in dermis. In conclusion, we demonstrate that it is possible to maintain live skin tissue cultures for 10 days. The viability analysis showed no significant signs of cell death in wound and suture models. The gene expression analysis demonstrated the interplay of activators and inhibitors of apoptosis and extracellular matrix components, thereby describing important features in ex vivo sutured wound healing models. Collectively, the performed methods defining analytical protocols proved to be applicable for post-flight analyzes of tissue specimens after sample return.
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http://dx.doi.org/10.3390/ijms18122604DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5751207PMC
December 2017

Photobiomodulation therapy by NIR laser in persistent pain: an analytical study in the rat.

Lasers Med Sci 2017 Nov 28;32(8):1835-1846. Epub 2017 Jul 28.

ASA Campus Joint Laboratory, ASA Res. Division-Department of Experimental and Clinical Biomedical Sciences "Mario Serio", University of Florence, Florence, Italy.

Over the past three decades, physicians have used laser sources for the management of different pain conditions obtaining controversial results that call for further investigations. In order to evaluate the pain relieving possibilities of photobiomodulation therapy (PBMT), we tested two near infrared (NIR) laser systems, with different power, against various kinds of persistent hyperalgesia animal models. In rats, articular pain was reproduced by the intra-articular injection of sodium monoiodoacetate (MIA) and complete Freund's adjuvant (CFA), while compressive neuropathy was modelled by the chronic constriction injury of the sciatic nerve (CCI). In MIA and CFA models, (NIR) laser (MLS-Mphi, ASA S.r.l., Vicenza, Italy) application was started 14 days after injury and was performed once a day for a total of 13 applications. In MIA-treated animals, the anti-hyperalgesic effect of laser began 5 min after treatment and vanished after 60 min. The subsequent applications evoked similar effects. In CFA-treated rats, laser efficacy started 5 min after treatment and disappeared after 180 min. In rats that underwent CCI, two treatment protocols with similar fluence but different power output were tested using a new experimental device called Multiwave Locked System laser (MLS-HPP). Treatments began 7 days after injury and were performed during 3 weeks for a total of 10 applications. Both protocols reduced mechanical hyperalgesia and hindlimb weight bearing alterations until 60 min after treatment with a higher efficacy recorded for the animals treated using the higher power output. In conclusion, this study supports laser therapy as a potential treatment for immediate relief of chronic articular or neuropathic pain.
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http://dx.doi.org/10.1007/s10103-017-2284-9DOI Listing
November 2017

A comparison trial between three treatment modalities for the management of myofascial pain of jaw muscles: A preliminary study.

Cranio 2018 Sep 11;36(5):327-331. Epub 2017 Jul 11.

c Department of Dentistry and Maxillofacial Surgery , Treviso Hospital , Treviso , Italy.

Objectives: To compare three treatment modalities for the management of myofascial pain of jaw muscles.

Methods: Thirty (N = 30) patients with low pain-related impairment were randomly assigned to receive laser therapy (LST), oral appliance therapy (OA), or counseling (CSL). Visual Analog Scale (VAS) pain levels and the Muscular Index (MI) of the Craniomandibular Index were the outcome variables, which were assessed at baseline, at three weeks, three months, and six months.

Results: At six months, improvement in the MI was maintained both in the LST (p = .025) and OA groups (p < .001). As for VAS values, positive changes were still shown for LST (p = .001), and were also shown for the OA (p = .002) and CSL groups (p = .048).

Conclusions: Despite differences in the short-term effectiveness of LST and OA, with respect to CSL alone, all three treatment groups improved at six months. This suggests that active treatments should be directed to maximize the positive changes in the short-term period.
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http://dx.doi.org/10.1080/08869634.2017.1349571DOI Listing
September 2018

Effects of extremely low-frequency magnetotherapy on proliferation of human dermal fibroblasts.

Electromagn Biol Med 2016 2;35(4):343-52. Epub 2016 Jun 2.

a Department of Biology and Biotechnologies "L. Spallanzani" , University of Pavia , Pavia , Italy.

Extremely low-frequency electromagnetic fields (ELF-EMFs) applied in magnetotherapy have frequency lower than 100 Hz and magnetic field intensity ranging from 0.1 to 20 mT. For many years, the use of magnetotherapy in clinics has been increasing because of its beneficial effects in many processes, e.g., skin diseases, inflammation and bone disorders. However, the understanding of the microscopic mechanisms governing such processes is still lacking and the results of the studies on the effects of ELF-EMFs are controversial because effects derive from different conditions and from intrinsic responsiveness of different cell types.In the present study, we studied the biological effects of 1.5 h exposure of human dermal fibroblasts to EMFs with frequencies of 5 and 50 Hz and intensity between 0.25 and 1.6 mT. Our data showed that the magnetic treatment did not produce changes in cell viability, but gave evidence of a sizeable decrease in proliferation at 24 h after treatment. In addition, immunofluorescence experiments displayed an increase in tubulin expression that could foreshadow changes in cell motility or morphology. The decrease in proliferation with unchanged viability and increase in tubulin expression could be consistent with the triggering of a transdifferentiation process after the exposure to ELF-EMFs.
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http://dx.doi.org/10.3109/15368378.2016.1138123DOI Listing
February 2017

In vitro study on the safety of near infrared laser therapy in its potential application as postmastectomy lymphedema treatment.

J Photochem Photobiol B 2015 Oct 11;151:285-96. Epub 2015 Aug 11.

ASAcampus Joint Laboratory, ASA Research Division, Department of Experimental and Clinical Biomedical Sciences "Mario Serio", University of Florence, Viale Pieraccini 6, I-50139 Florence, Italy. Electronic address:

Clinical studies demonstrated the effectiveness of laser therapy in the management of postmastectomy lymphedema, a discomforting disease that can arise after surgery/radiotherapy and gets progressively worse and chronic. However, safety issues restrict the possibility to treat cancer patients with laser therapy, since the effects of laser radiation on cancer cell behavior are not completely known and the possibility of activating postmastectomy residual cancer cells must be considered. This paper reports the results of an in vitro study aimed to investigate the effect of a class IV, dual-wavelength (808 nm and 905 nm), NIR laser system on the behavior of two human breast adenocarcinoma cell lines (namely, MCF7 and MDA-MB361 cell lines), using human dermal fibroblasts as normal control. Cell viability, proliferation, apoptosis, cell cycle and ability to form colonies were analyzed in order to perform a cell-based safety testing of the laser treatment in view of its potential application in the management of postmastectomy lymphedema. The results showed that, limited to the laser source, treatment conditions and experimental models used, laser radiation did not significantly affect the behavior of human breast adenocarcinoma cells, including their clonogenic efficiency. Although these results do not show any significant laser-induced modification of cancer cell behavior, further studies are needed to assess the possibility of safely applying NIR laser therapy for the management of postmastectomy lymphedema.
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http://dx.doi.org/10.1016/j.jphotobiol.2015.08.003DOI Listing
October 2015

Hypergravity stimulation enhances PC12 neuron-like cell differentiation.

Biomed Res Int 2015 16;2015:748121. Epub 2015 Feb 16.

Istituto Italiano di Tecnologia, Center for Micro-BioRobotics @SSSA, Viale Rinaldo Piaggio 34, 56025 Pontedera (Pisa), Italy.

Altered gravity is a strong physical cue able to elicit different cellular responses, representing a largely uninvestigated opportunity for tissue engineering/regenerative medicine applications. Our recent studies have shown that both proliferation and differentiation of C2C12 skeletal muscle cells can be enhanced by hypergravity treatment; given these results, PC12 neuron-like cells were chosen to test the hypothesis that hypergravity stimulation might also affect the behavior of neuronal cells, in particular promoting an enhanced differentiated phenotype. PC12 cells were thus cultured under differentiating conditions for either 12 h or 72 h before being stimulated with different values of hypergravity (50 g and 150 g). Effects of hypergravity were evaluated at transcriptional level 1 h and 48 h after the stimulation, and at protein level 48 h from hypergravity exposure, to assess its influence on neurite development over increasing differentiation times. PC12 differentiation resulted strongly affected by the hypergravity treatments; in particular, neurite length was significantly enhanced after exposure to high acceleration values. The achieved results suggest that hypergravity might induce a faster and higher neuronal differentiation and encourage further investigations on the potential of hypergravity in the preparation of cellular constructs for regenerative medicine and tissue engineering purposes.
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http://dx.doi.org/10.1155/2015/748121DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4345237PMC
November 2015

How microgravity affects the biology of living systems.

Biomed Res Int 2015 15;2015:863075. Epub 2015 Jan 15.

Department of Oral and Maxillofacial Surgery/Oral Pathology, VU-University Medical Center, 1081 HZ Amsterdam, Netherlands.

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http://dx.doi.org/10.1155/2015/863075DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4312564PMC
September 2015

The impact of microgravity and hypergravity on endothelial cells.

Biomed Res Int 2015 13;2015:434803. Epub 2015 Jan 13.

Department of Life Sciences, University of Siena, Via A. Moro 2, 53100 Siena, Italy.

The endothelial cells (ECs), which line the inner surface of vessels, play a fundamental role in maintaining vascular integrity and tissue homeostasis, since they regulate local blood flow and other physiological processes. ECs are highly sensitive to mechanical stress, including hypergravity and microgravity. Indeed, they undergo morphological and functional changes in response to alterations of gravity. In particular microgravity leads to changes in the production and expression of vasoactive and inflammatory mediators and adhesion molecules, which mainly result from changes in the remodelling of the cytoskeleton and the distribution of caveolae. These molecular modifications finely control cell survival, proliferation, apoptosis, migration, and angiogenesis. This review summarizes the state of the art on how microgravity and hypergravity affect cultured ECs functions and discusses some controversial issues reported in the literature.
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http://dx.doi.org/10.1155/2015/434803DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4309246PMC
October 2015

Effect of IR laser on myoblasts: a proteomic study.

Mol Biosyst 2013 Jun;9(6):1147-61

ASAcampus Joint Laboratory, ASA Res. Div., Dept. Clinical Physiopathology, University of Florence, Italy.

Laser therapy is used in physical medicine and rehabilitation to accelerate muscle recovery and in sports medicine to prevent damages produced by metabolic disturbances and inflammatory reactions after heavy exercise. The aim of this research was to get insight into possible benefits deriving from the application of an advanced IR laser system to counteract deficits of muscle energy metabolism and stimulate the recovery of hypotrophic tissue. We studied the effect of IR laser treatment on proliferation, differentiation, cytoskeleton organization and global protein expression in C2C12 myoblasts. We found that laser treatment induced a decrease in the cell proliferation rate without affecting cell viability, while leading to cytoskeletal rearrangement and expression of the early differentiation marker MyoD. The differential proteome analysis revealed the up-regulation and/or modulation of many proteins known to be involved in cell cycle regulation, cytoskeleton organization and differentiation.
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http://dx.doi.org/10.1039/c2mb25398dDOI Listing
June 2013

Hypergravity effects on myoblast proliferation and differentiation.

J Biosci Bioeng 2012 Feb 4;113(2):258-61. Epub 2011 Nov 4.

Italian Institute of Technology, Center of MicroBioRobotics c/o Scuola Superiore Sant'Anna, Viale Rinaldo Piaggio 34, 56025 Pontedera (Pisa), Italy.

This study aimed at the investigation of behavior of myoblasts in conditions of altered gravity. C2C12 cells underwent stimulations by different hypergravity intensities (2 h at 5 g, 10 g, and 20 g) in the Large Diameter Centrifuge of the European Space Agency (ESA), highlighting positive effects on both proliferation and differentiation.
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http://dx.doi.org/10.1016/j.jbiosc.2011.09.025DOI Listing
February 2012

Colorimetric comparison of light-filtering intraocular lenses and human crystalline lenses at various ages.

J Cataract Refract Surg 2011 Apr;37(4):758-62

Department of Clinical Physiopathology, University of Florence, Florence, Italy.

Purpose: To perform colorimetric characterization of 2 blue light-filtering intraocular lens (IOL) models and an IOL that filters ultraviolet-violet (UV-v) light and compare the charactizations with those of a standard UV light-filtering IOL and the human crystalline lens at various ages.

Setting: Department of Clinical Physiopathology, University of Florence, and National Institute of Applied Optics, CNR, Florence, Italy.

Design: Experimental study.

Methods: Three IOLs of each model with different dioptric powers (16.0, 21.0, and 26.0 diopters) were analyzed. The colorimetric coordinates of the IOLs were calculated by measuring their transmission spectra using a microspectroscopy technique. Subsequently, a parameter (ΔE) that was proportional to the perceived color differences was defined. The results of the IOLs and of the human crystalline lens at different ages (4, 22, 41, 53, and 72 years; values taken from the literature) were compared. The colorimetric analysis was performed using Commission Internationale de l'Éclairage standards.

Results: The maximum ΔE value was with the standard UV light-filtering IOL at any crystalline lens age and was greatest with a 72-year-old human lens. The minimum ΔE value was obtained for both blue light-filtering IOLs with middle-aged human lenses.

Conclusions: None of the analyzed IOLs had the same colorimetric performance of the human crystalline lens. The 2 blue light-filtering IOLs best reproduced yellowing of the human lens at 40 to 50 years of age.

Financial Disclosure: No author has a financial or proprietary interest in any material or method mentioned.
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http://dx.doi.org/10.1016/j.jcrs.2010.10.053DOI Listing
April 2011

Dendritic cells with lymphocyte-stimulating activity differentiate from human CD133 positive precursors.

Blood 2011 Apr 8;117(15):3983-95. Epub 2011 Feb 8.

Department of Anatomy, Histology and Forensic Medicine, University of Florence, Florence, Italy.

CD133 is a hallmark of primitive myeloid progenitors. We have addressed whether human cord blood cells selected for CD133 can generate dendritic cells, and Langerhans cells in particular, in conditions that promote that generation from CD34(+) progenitors. Transforming growth factor-β1 (TGF-β1) and anti-TGF-β1 antibody, respectively, were added in some experiments. With TGF-β, monocytoid cells were recognized after 7 days. Immunophenotypically immature dendritic cells were present at day 14. After 4 more days, the cells expressed CD54, CD80, CD83, and CD86 and were potent stimulators in mixed lymphocyte reaction; part of the cells expressed CD1a and langerin, but not Birbeck granules. Without TGF-β, only a small fraction of cells acquired a dendritic shape and expressed the maturation-related antigens, and lymphocytes were poorly stimulated. With anti-TGF-β, the cell growth was greatly hampered, CD54 and langerin were never expressed, and lymphocytes were stimulated weakly. In conclusion, CD133(+) progenitors can give rise in vitro, through definite steps, to mature, immunostimulatory dendritic cells with molecular features of Langerhans cells, although without Birbeck granules. Addition of TGF-β1 helps to stimulate cell growth and promotes the acquisition of mature immunophenotypical and functional features. Neither langerin nor Birbeck granules proved indispensable for lymphocyte stimulation.
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http://dx.doi.org/10.1182/blood-2010-08-299735DOI Listing
April 2011

Differentiating effects of the glucagon-like peptide-1 analogue exendin-4 in a human neuronal cell model.

Cell Mol Life Sci 2010 Nov 23;67(21):3711-23. Epub 2010 May 23.

Department of Clinical Physiopathology, Center for Research, Transfer and High Education on Chronic, Inflammatory, Degenerative and Neoplastic Disorders for the Development of Novel Therapies (DENOThe), University of Florence, Florence, Italy.

Glucagon-like peptide-1 (GLP-1) is an insulinotropic peptide with neurotrophic properties, as assessed in animal cell models. Exendin-4, a GLP-1 analogue, has been recently approved for the treatment of type 2 diabetes mellitus. The aim of this study was to morphologically, structurally, and functionally characterize the differentiating actions of exendin-4 using a human neuronal cell model (i.e., SH-SY5Y cells). We found that exendin-4 increased the number of neurites paralleled by dramatic changes in intracellular actin and tubulin distribution. Electrophysiological analyses showed an increase in cell membrane surface and in stretch-activated-channels sensitivity, an increased conductance of Na(+) channels and amplitude of Ca(++) currents (T- and L-type), typical of a more mature neuronal phenotype. To our knowledge, this is the first demonstration that exendin-4 promotes neuronal differentiation in human cells. Noteworthy, our data support the claimed favorable role of exendin-4 against diabetic neuropathy as well as against different neurodegenerative diseases.
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http://dx.doi.org/10.1007/s00018-010-0398-3DOI Listing
November 2010

Fibroblast autofluorescence in connective tissue disorders: a future tool for clinical and differential diagnosis?

J Biomed Opt 2008 Sep-Oct;13(5):054025

University of Florence, Department of Clinical Physiopathology, ASAcampus, ASA Research Division, Florence, Italy.

Marfan syndrome (MFS) is an inherited disorder of connective tissue due to mutations in FBN1 (90%) and TGFBR1 and TGFBR2 (5 to 10%) genes. Clinical and differential diagnosis is difficult because of the inter- and intrafamiliar marked heterogeneity and the variable onset age of clinical manifestations. Among the disorders, in differential diagnosis, thoracic aortic aneurysm (TAA) and Ullrich scleroatonic muscular dystrophy (UCMD) are reported. We evaluate the possibility of utilizing autofluorescence (AF) analysis as a diagnostic tool in the clinical and/or differential diagnosis of MFS and related disorders and in the investigation of the molecular mechanisms involved. Both multispectral imaging autofluorescence microscopy (MIAM) and autofluorescence microspectroscopy (AMS) have been used to characterize AF emission of fibroblasts from patients affected by inherited connective tissue disorders. Our preliminary results show significant differences in AF emission between normal and pathological fibroblasts, suggesting possible improvement in diagnostics of connective tissue disorders by AF analysis.
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http://dx.doi.org/10.1117/1.2982533DOI Listing
February 2009

A new method based on contact surface profilometry for quantitative measurement of resorbed bone volume.

Phys Med 2005 January - March;21(1):41-6

Dept. of Clinical Physiopathology, Medical Physics Sect. University of Florence, Viale Pieraccini 6, I 50139 Florence (Italy).

Bone is a dynamic tissue. Its continuous remodeling depends on the balance between bone formation and bone resorption. These two processes are carried out by specialized cells called osteoblast and ostreoclast respectively. The osteoclastic bone resorption consists in degradation of the mineral and collagen components of bone. The study of bone turnover requires accurate assessment of osteoclastic bone resorption, that becomes even more important in pathologic bone loss due to the uncoupling between bone formation and bone resorption. Osteoclastic activity is diffucult to measure. Many techniques, generally based on the detection of the resorbing lacunae (lacunae (pits) due to the bone degradation, allow to estimate bone resorption, but none of them quantitatively and directly measures the volume of resorbed bone. We propose a reliable and relatively simple method, based on contact surfact profilometry, to evaluate directly and quantitatively the volume of resorbed bone. The method has the following advantages:
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http://dx.doi.org/10.1016/S1120-1797(05)80018-3DOI Listing
October 2012

Multispectral imaging autofluorescence microscopy in colonic and gastric cancer metastatic lymph nodes.

Clin Gastroenterol Hepatol 2007 Feb;5(2):230-6

Department of Critical Medicine and Surgery, University of Florence, Florence, Italy.

Background & Aims: The lymphadenectomy and extended lymphadenectomy procedures have been points of controversy in surgical oncology. The methods available for the detection of metastatic lymph nodes are numerous. These include lymphoscintigraphy and radiolabeled antibody detection, but in most cancers the currently used technique is sentinel lymph node identification, performed primarily through the use of immunohistochemistry. We propose the application of autofluorescence (AF)-based techniques for lymph node evaluation in colorectal and gastric tumors.

Methods: We studied 30 clinical cases: 15 colorectal cancers and 15 gastric cancers. All of the patients were in the advanced stages of the disease and were candidates for adjuvant therapy. Autofluorescence microspectroscopy and multispectral imaging autofluorescence microscopy have been used to analyze the AF emission of metastatic lymph node sections, excited with 365-nm wavelength radiation. The AF spectra were recorded in the range of 400-700 nm. Monochrome AF images were acquired sequentially through interference filters peaked at 450, 550, and 650 nm, and then combined together in a single red-green-blue image. The AF pattern and the emission spectrum of metastatic lymph nodes have unique characteristics that can be used to distinguish them from the normal ones.

Results: The results, compared with standard histopathologic procedures and with specific staining methods, supplied a satisfactory validation of the proposed technique, revealing the possibility of improving the actual diagnostic procedures for malignant lymph node alterations.

Conclusions: With the development of appropriate instrumentation, the proposed technique could be particularly suitable in intrasurgical diagnosis of metastatic lymph nodes.
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http://dx.doi.org/10.1016/j.cgh.2006.11.013DOI Listing
February 2007

AcrySof Natural intraocular lens optical characteristics during and after different doses of ultraviolet-visible light illumination.

J Cataract Refract Surg 2006 Nov;32(11):1961-5

Department of Oto-Neuro-Ophthalmological Surgical Sciences-Eye Clinic, University of Florence, Italy.

Purpose: To determine the optical transmission properties of a recently introduced intraocular lens (IOL) (AcrySof Natural SN60AT, Alcon Laboratories) and compare them with those of a conventional IOL (AcrySof SA60AT, Alcon Laboratories) after ultraviolet (UV)-visible light irradiation.

Setting: Eye Clinic and Department of Clinical Physiopathology, University of Florence, and the National Institute of Applied Optics, Florence, Italy.

Methods: Two IOLs, the AcrySof Natural with a UV and blue-light filter and the AcrySof without a blue-light filter, were studied. Three IOLs of each model with different dioptric powers were examined in two experimental setups. The samples were stressed with very high irradiance to reproduce light aging, and the transmission properties of the IOLs were studied.

Results: No significant decrease in transmission was found at a UV-visible light dose equivalent to 100 years of light aging.

Conclusion: The AcrySof Natural IOL has stable optical characteristics.
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http://dx.doi.org/10.1016/j.jcrs.2006.06.024DOI Listing
November 2006

Modeled gravitational unloading triggers differentiation and apoptosis in preosteoclastic cells.

J Cell Biochem 2006 May;98(1):65-80

CEO--Center of Excellence in Optronics, L. Enrico Fermi 6, I-50125 Florence, Italy.

Gravity acts permanently on organisms as either static or dynamic stimulation. Understanding the influence of gravitational and mechanical stimuli on biological systems is an intriguing scientific problem. More than two decades of life science studies in low g, either real or modeled by clinostats, as well as experimentation with devices simulating different types of controlled mechanical stimuli, have shown that important biological functions are altered at the single cell level. Here, we show that the human leukemic line FLG 29.1, characterized as an osteoclastic precursor model, is directly sensitive to gravitational unloading, modeled by a random positioning machine (RPM). The phenotypic expression of cytoskeletal proteins, osteoclastic markers, and factors regulating apoptosis was investigated using histochemical and immunohistochemical methods, while the expression of the corresponding genes was analyzed using RT-PCR. A quantitative bone resorption assay was performed. Autofluorescence spectroscopy and imaging were applied to gain information on cell metabolism. The results show that modeled hypogravity may trigger both differentiation and apoptosis in FLG 29.1 cells. Indeed, when comparing RPM versus 1 x g cultures, in the former we found cytoskeletal alterations and a marked increase in apoptosis, but the surviving cells showed an osteoclastic-like morphology, overexpression of osteoclastic markers and the ability to resorb bone. In particular, the overexpression of both RANK and its ligand RANKL, maintained even after return to 1 x g conditions, is consistent with the firing of a differentiation process via a paracrine/autocrine mechanism.
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http://dx.doi.org/10.1002/jcb.20747DOI Listing
May 2006

Cell and tissue autofluorescence research and diagnostic applications.

Authors:
Monica Monici

Biotechnol Annu Rev 2005 ;11:227-56

Centre of Excellence in Optronics, Florence, Italy.

Cells contain molecules, which become fluorescent when excited by UV/Vis radiation of suitable wavelength. This fluorescence emission, arising from endogenous fluorophores, is an intrinsic property of cells and is called auto-fluorescence to be distinguished from fluorescent signals obtained by adding exogenous markers. The majority of cell auto-fluorescence originates from mitochondria and lysosomes. Together with aromatic amino acids and lipo-pigments, the most important endogenous fluorophores are pyridinic (NADPH) and flavin coenzymes. In tissues, the extracellular matrix often contributes to the auto-fluorescence emission more than the cellular component, because collagen and elastin have, among the endogenous fluorophores, a relatively high quantum yield. Changes occurring in the cell and tissue state during physiological and/or pathological processes result in modifications of the amount and distribution of endogenous fluorophores and chemical-physical properties of their microenvironment. Therefore, analytical techniques based on auto-fluorescence monitoring can be utilized in order to obtain information about morphological and physiological state of cells and tissues. Moreover, auto-fluorescence analysis can be performed in real time because it does not require any treatment of fixing or staining of the specimens. In the past few years spectroscopic and imaging techniques have been developed for many different applications both in basic research and diagnostics.
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http://dx.doi.org/10.1016/S1387-2656(05)11007-2DOI Listing
July 2006

Simulated hypogravity impairs the angiogenic response of endothelium by up-regulating apoptotic signals.

Biochem Biophys Res Commun 2005 Aug;334(2):491-9

Section of Pharmacology, Department of Molecular Biology and C.R.I.S.M.A., University of Siena, Siena, Italy.

Health hazards in astronauts are represented by cardiovascular problems and impaired bone healing. These disturbances are characterized by a common event, the loss of function by vascular endothelium, leading to impaired angiogenesis. We investigated whether the exposure of cultured endothelial cells to hypogravity condition could affect their behaviour in terms of functional activity, biochemical responses, morphology, and gene expression. Simulated hypogravity conditions for 72 h produced a reduction of cell number. Genomic analysis of endothelial cells exposed to hypogravity revealed that proapoptotic signals increased, while antiapoptotic and proliferation/survival genes were down-regulated by modelled low gravity. Activation of apoptosis was accompanied by morphological changes with mitochondrial disassembly and organelles/cytoplasmic NAD(P)H redistribution, as evidenced by autofluorescence analysis. In this condition cells were not able to respond to angiogenic stimuli in terms of migration and proliferation. Our study documents functional, morphological, and transcription alterations in vascular endothelium exposed to simulated low gravity conditions, thus providing insights on the occurrence of vascular tissue dysregulation in crewmen during prolonged space flights. Moreover, the alteration of vascular endothelium can intervene as a concause in other systemic effects, like bone remodelling, observed in weightlessness.
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http://dx.doi.org/10.1016/j.bbrc.2005.06.124DOI Listing
August 2005

Dependence of leukemic cell autofluorescence patterns on the degree of differentiation.

Photochem Photobiol Sci 2003 Oct;2(10):981-7

Centre of Excellence in Optronics, Largo Enrico Fermi, 6, 50125 Florence, Italy.

The characterisation of leukemic cell autofluorescence during differentiation, induced by 12-O-tetradecanoylphorbol 13-acetate and all-trans retinoic acid, was performed by autofluorescence microspectroscopy and multispectral imaging autofluorescence microscopy. We have found that a dependence exists between the cell autofluorescence pattern and the degree of cell differentiation. When cells differentiate, their autofluorescence emission changes, following the morphological and functional rearrangement of cell structures. A decrease in emission intensity and a different distribution of endogenous fluorophores are observed. Thus, autofluorescence monitoring on living cells is a potentially useful tool for in vitro study of the differentiation processes. Furthermore, different maturation steps can be distinguished on the basis of the cell fluorescence pattern, leading the way to future application of the technique in diagnostics.
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http://dx.doi.org/10.1039/b306276gDOI Listing
October 2003