Bone marrow-derived mesenchymal stem cells attenuate pulmonary inflammation and lung damage caused by highly pathogenic avian influenza A/H5N1 virus in BALB/c mice.

BACKGROUND: The highly pathogenic avian influenza A/H5N1 virus is one of the causative agents of acute lung injury (ALI) with high mortality rate. Studies on therapeutic administration of bone marrow-derived mesenchymal stem cells (MSCs) in ALI caused by the viral infection have been limited in number and have shown conflicting results. The aim of the present investigation is to evaluate the therapeutic potential of MSC administration in A/H5N1-caused ALI, using a mouse model. METHODS: MSCs were prepared from the bone marrow of 9 to 12 week-old BALB/c mice. An H5N1 virus of A/turkey/East Java/Av154/2013 was intranasally inoculated into BALB/c mice. On days 2, 4, and 6 after virus inoculation, MSCs were intravenously administered into the mice. To evaluate effects of the treatment, we examined for lung alveolar protein as an indicator for lung injury, PaO2/FiO2 ratio for lung functioning, and lung histopathology. Expressions of NF-κB, RAGE (transmembrane receptor for damage associated molecular patterns), TNFα, IL-1ß, Sftpc (alveolar cell type II marker), and Aqp5+ (alveolar cell type I marker) were examined by immunohistochemistry. In addition, body weight, virus growth in lung and brain, and duration of survival were measured. RESULTS: The administration of MSCs lowered the level of lung damage in the virus-infected mice, as shown by measuring lung alveolar protein, PaO2/FiO2 ratio, and histopathological score. In the MSC-treated group, the expressions of NF-κB, RAGE, TNFα, and IL-1ß were significantly suppressed in comparison with a mock-treated group, while those of Sftpc and Aqp5+ were enhanced. Body weight, virus growth, and survival period were not significantly different between the groups. CONCLUSION: The administration of MSCs prevented further lung injury and inflammation, and enhanced alveolar cell type II and I regeneration, while it did not significantly affect viral proliferation and mouse morbidity and mortality. The results suggested that MSC administration was a promissing strategy for treatment of acute lung injuries caused by the highly pathogenic avian influenza A/H5N1 virus, although further optimization and combination use of anti-viral drugs will be obviously required to achieve the goal of reducing mortality.

Endemic and emerging acute virus infections in Indonesia: an overview of the past decade and implications for the future.

Being the largest archipelago country in the world, with a tropical climate and a unique flora and fauna, Indonesia habitats one of the most diverse biome in the world. These characteristics make Indonesia a popular travel destination, with tourism numbers increasing yearly. These characteristics also facilitate the transmission of zoonosis and provide ideal living and breading circumstances for arthropods, known vectors for viral diseases. A review of the past 10 years of literature, reports of the Ministry of Health, Republic of Indonesia and ProMED-mail shows a significant increase in dengue infection incidence. Furthermore, chikungunya, Japanese encephalitis and rabies are proven to be endemic in Indonesia. The combination of cohort studies, governmental data and ProMED-mail reveals an integrated overview for those working in travel medicine and public health, focusing on both endemic and emerging acute virus infections. This review summarizes the epidemiology of acute virus infections in Indonesia, including outbreak reports, as well as public health response measurements and their potential or efficacy. Knowledge about human behaviour, animal reservoirs, climate factors, environment and their role in emerging virus infection are discussed. We aim to support public health authorities and health care policy makers in a One Health approach.

The optimum oxygen level in hypoxic culture conditions of ligament derived stem cells: experimental research.

The hypoxic condition is a physiological norm for various stem cells. The natural microenvironment contains lower oxygen pressures. Recent studies reported significant increases in the cultured cells' proliferation in the presence of a low oxygen pressure. Objective: This study aimed to investigate the optimum oxygen level for rabbit cruciate ligament fibroblast cells culture and Ligament Derived Conditioned Medium/LD-CM (Secretome) preparation in vitro. Materials and methods: Fibroblasts were isolated from the cruciate ligament of the rabbit's knee. Cultured of rabbit cruciate ligament Fibroblast Cells (fifth passage) were assigned to the slight (5% O2), middle (3% O2), and severe hypoxia (1% O2) groups and the normoxia (21% O2) group. Measurement of growth factors: TGF-ß1, PDGF, FGF, and VEGF in LD-CM (Secretome) used an enzyme-linked immunosorbent assay. Results: The highest number of cultured cells were in the 5% O2 group compared to the normoxia, 1 and 3% groups. The hypoxia 5% group also had increased productions of PDGF, FGF, and VEGF proteins in LD-CM (secretome) compared to the 1, 3%, and normoxia groups. TGF-ß1 production was slightly higher in the 3 group than the 5% group. Conclusion: The hypoxic precondition of 5% oxygen was the optimum condition for ligament culture and ligament derived conditioned medium (secretome) preparation in vitro.

The Comparison between Microneedling and Fractional CO2 Laser for Amniotic Membrane Stem Cell-Conditioned Medium and Vitamin C in Photoaging Treatment.

Context: Photoaging correlates with ultraviolet radiation (UVR). In addition to direct and indirect DNA damage, UVR leads to a breakdown of collagen, resulting in enlarged pores, visible wrinkles, also uneven skin tones and pigmentations. Amniotic membrane stem cell (AMSC) can be a useful modality in cell regeneration. Vitamin C has been known as an agent of depigmentation, antioxidants, and collagen synthesis. A topical combination of amniotic membrane stem cell-conditioned medium (AMSC-CM) and vitamin C is expected to improve photoaging. Fractional CO2 laser and microneedling served as transdermal drug delivery. Aims: To compare between microneedling and fractional CO2 laser for the administration of AMSC-CM and vitamin C in photoaging treatment. Methods and Material: Sixty Indonesian women with photoaged skins participated in this experimental analytic, controlled, matching research and were divided into two groups. Thirty women received fractional CO2 laser, and others received microneedling. All participants received a topical combination of AMSC-CM and vitamin C. Three treatment sessions were repeated at a four-week interval for fractional CO2 laser and a two-week interval for microneedling. We evaluated the progression and side effects using a skin analyzer on weeks 0, 4, and 8 for the microneedling group and for the fractional CO2 laser group on weeks 0, 4, and 12. Results: Assessment of wrinkles, UV spots, skin tone, and pore revealed a statistically significant effect of a topical combination of AMSC-CM and vitamin C after fractional CO2 laser (p < 0.05). Conclusions: Combination of AMSC-CM and vitamin c using fractional CO2 laser can improve photoaging and is better compared to microneedling.

Resveratrol promotes secretion of wound healing related growth factors of mesenchymal stem cells originated from adult and fetal tissues.

Non-healing wounds have demonstrated aberrant regulation of several growth factors, thus using exogenous growth factors and cytokines in the clinical setting may improve the outcomes of non-healing wounds. Mesenchymal stem cells (MSCs) are the source of growth factors that show beneficial effect in promoting impaired wound healing. Certain culture condition should be developed to stimulate growth factor secretion from stem cell. Resveratrol, a small molecule found to increase MSCs therapeutic effectiveness. This study aims to investigate the effect of RV on secretion of wound healing related growth factors. We isolated and characterised MSCs from wharton's jelly (WJ), amniotic membrane (AM), and adipose tissue. We treated MSCs with serum deprived medium (SDM) supplemented with RV at 0.1 µM, 0.5 µM, 0.8 µM concentration. Our study revealed that RV at 0.1 µM was more effective to increase cell proliferation rate. Resveratrol at 0.1 µM promoted EGF, HGF, PDGF, and TGF-ß1 secretion from MSCs. AD-MSCs showed the greatest response to RV stimulation in the term of cell proliferation and growth factors secretion. As conclusion, RV can facilitate cell proliferation and wound healing related growth factors secretion at dosage dependent manner.

Comparison of Virulence and Lethality in Mice for Avian Influenza Viruses of Two A/H5N1 and One A/H3N6 Isolated from Poultry during Year 2013-2014 in Indonesia.

In Indonesia, the highly pathogenic avian influenza A/H5N1 virus has become endemic and has been linked with direct transmission to humans. From 2013 to 2014, we isolated avian influenza A/H5N1 and A/H3N6 viruses from poultry in Indonesia. This study aimed to reveal their pathogenicity in mammals using a mouse model. Three of the isolates, Av154 of A/H5N1 clade 2.3.2.1c, Av240 of A/H5N1 clade 2.1.3.2b, and Av39 of A/H3N6, were inoculated into BALB/c mice. To assess morbidity and mortality, we measured body weight daily and monitored survival for 20 d. Av154- and Av240-infected mice lost 25% of their starting body weight by day 7, while Av39-infected mice did not. Most of the Av154-infected mice died on day 8, while the majority of the Av240-infected mice survived until day 20. A 50% mouse lethal dose was calculated to be 2.0 × 101 50% egg infectious doses for Av154, 1.1 × 105 for Av240 and > 3.2 × 106 for Av39. The Av154 virus was highly virulent and lethal in mice without prior adaptation, suggesting its high pathogenic potential in mammals. The Av240 virus was highly virulent but modestly lethal, whereas the Av39 virus was neither virulent nor lethal. Several mammalian adaptive markers of amino acid residues were associated with the highly virulent and lethal phenotypes of the Av154 virus.

The effects of amniotic membrane stem cell-conditioned medium on photoaging.

Background: Photoaging is a complex biologic process that affects various layers of the skin with the major damage seen in the connective tissue of the dermis. Although rarely fatal, photoaging can significantly contribute to a loss of quality of life; therefore, it deserves attention. Researchers are continually searching for new compounds to develop rejuvenation therapies. Nowadays, the stem cell population has been discovered, and it shows capability to rejuvenate the skin. Material and methods: This was an analytic experimental research conducted on a total of 48 women who were randomized into two groups: 24 women received amniotic membrane stem cell-conditioned medium (AMSC-CM) and the other 24 women received normal saline (NS). We applied the drug for three times with an interval of 2 weeks. Microneedling was used to enhance epidermal penetration. We evaluated the progression of photoaging on Weeks 0, 4, and 8, as well as the side effects. Results: The improvement in photoaging after treatment showed significant better effects with the AMSC-CM than with NS (p < .05). There were minimal transient side effects in either of the study groups. Conclusion: AMSC-CM has the capability to improve clinical photoaging and is a promising option for rejuvenation therapy.

Biocompatibility of Yttria-Tetragonal Zirconia Polycrystal Seeded with Human Adipose Derived Mesenchymal Stem Cell.

INTRODUCTION: The scaffold is a place for regeneration of new bone and bone tissue growths in tissue engineering applications. hADMSC is a multipotent cell which can differentiate into osteogenic, chondrogenic and adipogenic. Y-TZP has been shown to have several advantages over other ceramics because of its hard nature, namely fracture toughness and high flexural strength. AIM: This study aimed to analyze the biocompatibility of Y-TZP as a scaffold seeded with hADMSCs by in vitro analysis. MATERIAL AND METHODS: This research involved several processes, namely Y-TZPS manufacture process, XRD examination, differentiation and characterization of hADMSC, SEM observation, and then TT. RESULTS: The results of the XRD examination showed that Y-TZPSs had sharp peaks. It suggests that they had high crystal purity. The marked expression of the characterization of hADMSC is the positive expression of Cluster of differentiation (CD), namely CD 90, CD 73 and CD 105 above NMT and negative expressions of CD 14, CD 19, CD 34, CD 45 and also HLA-DR below NLT. The analysis of observations on the Y-TZPSs with SEM, subsequently, indicated the porosity of Y-TZPSs, as a result, the adhesion of HADMSCs occurred and grew in the porosity in the Y-TZPSs. CONCLUSIONS: Y-TZPSs with low porosity and toxicity can be able to proliferate and differentiate if seeded with hADMSC. Y-TZPSs are expected to be used as implantable biomaterials using hADMSCs with high biocompatibility.

Effect of Allogeneic Bone Marrow-mesenchymal Stem Cells (BM-MSCs) to Accelerate Burn Healing of Rat on the Expression of Collagen Type I and Integrin α2ß1.

BACKGROUND AND OBJECTIVE: Burn is a public health problem, it causes physical disability even death. Treatment of burn wound has been conducted in various ways, but the satisfactory healing has not been provided. Bone marrow-derived mesenchymal stem cells (BM-MSCs) treatment is one of attempt to burn recovery, accelerate wound healing and angiogenesis. This study aimed to investigate the effect of allogeneic BM-MSC treatment on the expression of collagen type I and integrin α2ß1 in burn skin tissue of rat observed on day 14. MATERIALS AND METHODS: Twelve Wistar rats divided into two groups, control group (injected with phospate buffer solution) and treatment group (injected with BM-MSC). Rat was anaesthetized with xylazine and ketamine (ratio 1:1), fur of rat's back was shaved and full thickness burn was made by boiling plate in hot water for 30 min and patched on the back for 20 min. The burns were covered by tegaderm film and elastomult haft. Antalgin as an analgetic was injected to rats during observation process. Burns of rat was observed on day 14. In this study one-way analysis of variance test and Tukey as a further test were analyzed. RESULTS: The results showed that the healing time of allogeneic BM-MSC treatment on burn skin tissue rats was faster, the thickness of collagen type I in burn skin tissue of rats was thicker (0.977 µm) than controls (0.475 µm) and statistically demonstrated significant differences (p = 0.000). The average percentage of integrin α2ß1 expression was higher (2.94%) than control group (2.34%), but the differences were not statistically significant (p = 0.176). CONCLUSION: The study concluded that BM-MSC treatment was able to accelerate the healing process of burns by increasing the thickness of the collagen and the percentage of integrin α2ß1, thus accelerated the cell migration involved during wound healing.

Displacement of the predominant dengue virus from type 2 to type 1 with a subsequent genotype shift from IV to I in Surabaya, Indonesia 2008-2010.

Indonesia has annually experienced approximately 100,000 reported cases of dengue fever (DF) and dengue hemorrhagic fever (DHF) in recent years. However, epidemiological surveys of dengue viruses (DENVs) have been limited in this country. In Surabaya, the second largest city, a single report indicated that dengue virus type 2 (DENV2) was the predominant circulating virus in 2003-2005. We conducted three surveys in Surabaya during: (i) April 2007, (ii) June 2008 to April 2009, and (iii) September 2009 to December 2010. A total of 231 isolates were obtained from dengue patients and examined by PCR typing. We found that the predominant DENV shifted from type 2 to type 1 between October and November 2008. Another survey using wild-caught mosquitoes in April 2009 confirmed that dengue type 1 virus (DENV1) was the predominant type in Surabaya. Phylogenetic analyses of the nucleotide sequences of the complete envelope gene of DENV1 indicated that all 22 selected isolates in the second survey belonged to genotype IV and all 17 selected isolates in the third survey belonged to genotype I, indicating a genotype shift between April and September 2009. Furthermore, in December 2010, isolates were grouped into a new clade of DENV1 genotype I, suggesting clade shift between September and December 2010. According to statistics reported by the Surabaya Health Office, the proportion of DHF cases among the total number of dengue cases increased about three times after the type shift in 2008. In addition, the subsequent genotype shift in 2009 was associated with the increased number of total dengue cases. This indicates the need for continuous surveillance of circulating viruses to predict the risk of DHF and DF.