Isolation and Characterization of Skin Derived Mesenchymal Stem Cell (SMSCs) from New Zealand Rabbit, Oryctolagus Cuniculus: A in Vitro Study

Mesenchymal stem cells (MSCs) exhibit high proliferation and self-renewal capabilities and are critical for tissue repair and regeneration. Furthermore, they can be isolated from a variety of tissues and have many potential applications in the clinical setting. In this study, Skin Derived Mesenchymal Stem Cells (SMSCs) were isolated from a New Zealand rabbit and analyzed for the skin sampling area, separation method, culture conditions, primary and passage culture times, and cell surface markers. The result shows that SMSCs have surface markers of mesenchymal stem cells by immunofluorescence of CD105, CD90, CD73, CD44, and CD200 and the CD105, CD90, and CD73 positive markers were also confirmed with Flow cytometry. Furthermore, negative markers of CD45 and CD34 did not show in immunofluorescence but CD45, CD34, and CD133 were confirmed with Flow cytometry

A Comparative study on the therapeutic potential of ocular and non-ocular stem cell secretome on alkali induced limbal stem cell niche damage

The study of secretome from stem cells has become a promising area of research in limbal stem cell deficiency (LSCD). This study aimed to investigate the therapeutic potential of secretome from AD-MSCs and LSCs in a culture of alkalidamaged LSCs as a model of limbal niche damage. Secretome was derived from 4 passages of AD-MSCs and LSCs under serum deprivation conditions. LSCs were damaged using 6,25mM NaOH to generate a limbal niche damage model and were then divided into 5 groups consisting of a control group, as well as 50% AD-MSCs, 25% AD-MSCs, 50% LSCs and 25% LSCs as the treated groups. After 48 hours of treatment, cell proliferation was determined by MTT-assay and cell migration was determined by in vitro scratch assay. VEGF supernatant was measured to analyze anti-angiogenic properties. A concentration of EGF and HGF in secretome was measured using ELISA. The data were analyzed using Oneway ANOVA dan post-hoc Tukey. Secretome derived from AD-MSCs and LSCs successfully promoted the proliferation and migration of LSCs, compared to the control, with the highest proliferation and migration rate seen in the 50% LSC secretome group ((94.38±2.51%, P 0.000; 68.52 ± 3.51%, P 0.000, respectively). EGF (1216.67 pg/mL) and HGF (2943.33 pg/mL) levels were higher in LSC secretome compared to the secretome of AD-MSCs. A 50% AD-MSC secretome with a level of VEGF supernatant at 65 pg/mL showed better anti-angiogenic activity. Secretome from ocular and non-ocular stem cells demonstrated clear therapeutic benefits inlimbal niche damage

The potency of hematopoietic stem cells (hscs) and natural killer (nk) cells as a therapeutic of sars-cov-2 Indonesia isolates infection by viral inactivation (in vitro Study)

Background: The prevalence of COVID-19 cases in Indonesia as of June 9, 2020, has been confirmed 32.076 positive cases, with 1.923 death cases. The total number of deaths reached 92,941 cases. There has been a recent update on stem cell-based biological, medical therapy as an optional treatment to handling COVID-19 due to its potential viability besides using the prevalent conventional chemical drug therapy. Methods: In this study, in vitro research was conducted to determine the potential of hematopoietic stem cells (HSCs) and natural killer cells (NK cells) against SARS-CoV-2 viruses, which virus isolates were collected in Indonesia. The SARS-CoV-2 virus was planted in rat kidney cells and Vero cells. The cells that had been planted with the virus were given HSCs and NK cells, followed by being evaluated at intervals of 24, 48, and 72 hours. The evaluation was done by collecting cells and supernatant from the cell plate and then determining the viral load using a Polymerase Chain Reaction (PCR) machine. Results: The results showed that the addition of HSCs and NK on cells that had been infected by SARS-CoV-2 resulted in a decrease in viral load within 24 to 72 hours in all variations of Multiples of Infection (MoI) values. Conclusions: The administration of HSCs and NK cells has the potential to eliminate the SARS-CoV-2 virus. Although this study is only an in vitro study, it could be the basis for the development of alternative stem cell-based therapies to tackle COVID-19 cases

Gingival Mesenchymal Stem Cells from Wistar Rat’s Gingiva (Rattus Novergicus) – Isolation and Characterization (In Vitro Study)

Gingiva is emerging as a source of Mesenchymal Stem Cells. Gingival Mesenchymal Stem Cell has been isolated and characterized from the gingival connective tissue of wistar rat (Rattus Novergicus). Gingival Mesenchymal Stem Cell sources are rich, attainable and easy to isolate through minimal invasive procedure. Gingival Mesenchymal Stem Cells are ideal to accelerate bone regeneration. The aim of this study was to analyze Gingival Mesenchymal Stem Cells from Wistar Rats’ gingiva (Rattus Novergicus) isolation and characterization by CD34, CD44, CD73, CD90, CD105 expression. This study was descriptive observational with simple random sampling method. Gingival Mesenchymal Stem Cells were isolated from healthy, 200 gram, 1 month year old, male rat’s (Rattus Novergicus) lower gingival tissue through gingivectomy procedure (n=4). Gingiva were minced into small fragments then cultured in 2 weeks. The culture was passaged every 3-5 days after cultured and plated. The isolated Gingival Mesenchymal Stem Cells in passage 5 were characterized by CD34, CD44, CD73, CD90, CD105 using Immunocytochemistry and flowcytometry examination. Gingival Mesenchymal Stem Cells strongly expressed CD44+, CD73+, CD90+, CD105+ but did not express CD45- and CD34-. Gingival Mesenchymal Stem Cells’ morphology was fibroblast-like, spindle-shaped, colony-forming abilities, and stick to the culture plate. Gingiva is potential Stem Cell source. Gingival Mesenchymal Stem Cells has multipotency ability with proliferation and mesenchymal stem cells characteristic advantageous for tissue engineering and regenerative therapy

The Potency of Hematopoietic Stem Cells (HSCs) and Natural Killer (NK) Cells as A Therapeutic of SARS-CoV-2 Indonesia Isolates Infection by Viral Inactivation (In Vitro Study)

Background: The prevalence of COVID-19 cases in Indonesia as of June 9, 2020, has been confirmed 32.076 positive cases, with 1.923 death cases. The total number of deaths reached 92,941 cases. There has been a recent update on stem cell-based biological, medical therapy as an optional treatment to handling COVID-19 due to its potential viability besides using the prevalent conventional chemical drug therapy. Methods: In this study, in vitro research was conducted to determine the potential of hematopoietic stem cells (HSCs) and natural killer cells (NK cells) against SARS-CoV-2 viruses, which virus isolates were collected in Indonesia. The SARS-CoV-2 virus was planted in rat kidney cells and Vero cells. The cells that had been planted with the virus were given HSCs and NK cells, followed by being evaluated at intervals of 24, 48, and 72 hours. The evaluation was done by collecting cells and supernatant from the cell plate and then determining the viral load using a Polymerase Chain Reaction (PCR) machine. Results: The results showed that the addition of HSCs and NK on cells that had been infected by SARS-CoV-2 resulted in a decrease in viral load within 24 to 72 hours in all variations of Multiples of Infection (MoI) values. Conclusions: The administration of HSCs and NK cells has the potential to eliminate the SARS-CoV-2 virus. Although this study is only an in vitro study, it could be the basis for the development of alternative stem cell-based therapies to tackle COVID-19 cases

An in vitro study of dual drug combinations of anti-viral agents, antibiotics, and/or hydroxychloroquine against the SARS-CoV-2 virus isolated from hospitalized patients in Surabaya, Indonesia

A potent therapy for the infectious coronavirus disease COVID-19 is urgently required with, at the time of writing, research in this area still ongoing. This study aims to evaluate the in vitro anti-viral activities of combinations of certain commercially available drugs that have recently formed part of COVID-19 therapy. Dual combinatory drugs, namely; LopinavirRitonavir (LOPIRITO)-Clarithromycin (CLA), LOPIRITO-Azithromycin (AZI), LOPIRITODoxycycline (DOXY), Hydroxychloroquine (HCQ)-AZI, HCQ-DOXY, Favipiravir (FAVI)-AZI, HCQ-FAVI, and HCQ-LOPIRITO, were prepared. These drugs were mixed at specific ratios and evaluated for their safe use based on the cytotoxicity concentration (CC50) values of human umbilical cord mesenchymal stem cells. The anti-viral efficacy of these combinations in relation to Vero cells infected with SARS-CoV-2 virus isolated from a patient in Universitas Airlangga hospital, Surabaya, Indonesia and evaluated for IC50 24, 48, and 72 hours after viral inoculation was subsequently determined. Observation of the viral load in qRT-PCR was undertaken, the results of which indicated the absence of high levels of cytotoxicity in any samples and that dual combinatory drugs produced lower cytotoxicity than single drugs. In addition, these combinations demonstrated considerable effectiveness in reducing the copy number of the virus at 48 and 72 hours, while even at 24 hours, post-drug incubation resulted in low IC50 values. Most combination drugs reduced pro-inflammatory markers, i.e. IL-6 and TNF-α, while increasing the anti-inflammatory response of IL-10. According to these results, the descending order of effective dual combinatory drugs is one of LOPIRITO-AZI>LOPIRITO-DOXY>HCQ-AZI>HCQ FAVI>LOPIRITO-CLA>HCQ-DOX. It can be suggested that dual combinatory drugs, e.g. LOPIRITO-AZI, can potentially be used in the treatment of COVID-19 infectious diseases

Prevalensi Ektoparasit Protozoa Lchthyophthirius Multifiliis pada Ikan Maskoki (Carassius Auratus) di Desa Canggu Kecamatan Pare Kabupaten Kediri

This study aims were to know the prevalence of Ichthyophthirius multifiliis infection on goldfish fantail aquaculture, knowing the number of Ichthyophthirius multifiliis infection on age of fishes were 2 months and 4 months old, knowing the number of khthyophthirius multifiliis infection which infect skins and gills, and knowing the number of Ichthyophthirius multifiliis infection in two places of aquaculture. This study was conducted on March 2011 in Canggu village, Pare - Kediri. This study used 120 samples consists of two ages were 2 months and 4 months old. The method of examination used in this study was skin scraping and gill biopsy. The results showed that prevalence of Ichthyophthirius multifiliis in Canggu village, Pare - Kediri was 15.8%. The infection in the young fish was 9.2% and the older ones was 6.7%. The gill infection was 13.3% and the skin infection was 2.5%. Otherwise, the infection in the aquaculture A was 7.5% and in the aquaculture B was 8.3%

A Randomized, Double-Blind, Multicenter Clinical Study Comparing the Efficacy and Safety of a Drug Combination of Lopinavir/Ritonavir-Azithromycin, Lopinavir/Ritonavir-Doxycycline, and Azithromycin-Hydroxychloroquine for Patients Diagnosed with Mild to Moderate COVID-19 Infections

At the present time, COVID-19 vaccines are at the testing stage, and an effective treatment for COVID-19 incorporating appropriate safety measures remains the most significant obstacle to be overcome. A strategic countermeasure is, therefore, urgently required. Aim. +is study aims to evaluate the efficacy and safety of a combination of lopinavir/ritonavirazithromycin, lopinavir/ritonavir-doxycycline, and azithromycin-hydroxychloroquine used to treat patients with mild to moderate COVID-19 infections. Setting and Design. +is study was conducted at four different clinical study sites in Indonesia. +e subjects gave informed consent for their participation and were confirmed as being COVID-19-positive by means of an RTPCR test. +e present study constituted a randomized, double-blind, and multicenter clinical study of patients diagnosed with mild to moderate COVID-19 infection. Materials and Methods. Six treatment groups participated in this study: a Control group Madministered with a 500 mg dose of azithromycin; Group A which received a 200/50 mg dose of lopinavir/ritonavir and 500 mg of azithromycin; Group B treated with a 200/50 mg dose of lopinavir/ritonavir and 200 mg of doxycycline; Group C administered with 200 mg of hydroxychloroquine and 500 mg of azithromycin; Group D which received a 400/100 mg dose of lopinavir/ritonavir and 500 mg of azithromycin; and Group E treated with a 400/100 mg dose of lopinavir/ritonavir and 200 mg of doxycycline. Results. 754 subjects participated in this study: 694 patients (92.4%) who presented mild symptoms and 57 patients (7.6%) classified as suffering from a moderate case of COVID-19. On the third day after treatment, 91.7%–99.2% of the subjects in Groups A–E were confirmed negative by a PCR swab test compared to 26.9% in the Control group. Observation of all groups which experienced a significant decrease in virus load between day 1 and day 7 was undertaken. Other markers, such as CRP and IL-6, were significantly lower in all treatment groups (p< 0.05 and p< 0.0001) than in the Control group. Furthermore, IL-10 and TNF-α levels were significantly elevated in all treatment groups (p< 0.0001). +e administration of azithromycin to the Control group increased CRP and IL-6 levels, while reduced IL-10 and TNF-α on day 7 (p< 0.0001) compared with day 1. Decreases in ALTand AST levels were observed in all groups (p< 0.0001). +ere was an increase in creatinine in the serum level of the Control, C, D, and E groups (p< 0.05), whereas the BUN level was elevated in all groups (p< 0.0001). Conclusions. +e study findings suggest that the administration of lopinavir/ritonavir-doxycycline, lopinavir/ritonavir-azithromycin, and azithromycinhydroxychloroquine as a dual drug combination produced a significantly rapid PCR conversion rate to negative in three-day treatment of mild to moderate COVID-19 cases. Further studies should involve observation of older patients with severe clinical symptoms in order to collate significant amounts of demographic data

Metode Pembuatan Metabolit Sel Punca Dari Darah Tali Pusat Manusia Untuk Regenerasi Kulit

Invensi ini berkaitan dengan metode pemrosesan produk metabolit stem cell untuk regenerasi kulit yang dilakukan di Laboratorium. Etabolit stem cell yang dihasilkan berasal dari darah tali pusat bayi dari honor sukarela yang memenuhi standar, untuk kemudian dilakukan kultur dan diferensiasi menjadi stem cell mesenkim. Kandungan sitokin dalam metabolit stem cell yaitu IL-10, IL-4, EGF, GM-CSF dan TGF-B yang berperan dalam mempercepat regenerasi kulit, merangsang pertumbuhan dan perkembangan berbagai sel kekebalan, menggantikan sel-sel yang rusak, membuat kulit memproduksi sendiri serat elastic protein agar kulit kembali elastis, serta memproduksi lebih banyak kolagen dan elastin. Darah tali pusat diisolasi kemudian dikultur pada komplit medium dalam sistem bioreaktor. Selanjutnya dikarakterisasi menggunakan flow cytometry dengan marker psoitif CD105 dan CD90

The distribution pattern and growth factor level in platelet-rich fibrin incorporated skin-derived mesenchymal stem cells: An in vitro study

Background and Aim: A skin wound in an animal must be cared for to prevent further health issues. Platelet-rich fibrin (PRF) and skin-derived mesenchymal stem cells (SMSCs) have been reported to have potential in increasing the rate of wound healing. This study aimed to analyze the distribution patterns and levels of platelet-derived growth factor (PDGF), insulin-like growth factor (IGF), vascular endothelial growth factor (VEGF), and transforming growth factor-β (TGF-β) in PRF incorporated with SMSCs. Materials and Methods: This study employed a true experiment (in vitro) design with post-test only performed in the control group alone. PRF and SMSCs were extracted from the blood and skin of 16 rabbits. SMSCs were characterized using immunocytochemistry to examine clusters of differentiation for 45, 73, 90, and 105. PRF was incorporated into the SMSCs and then divided into four groups (N=32/n=8): Group A (PRF only), Group B (PRF+SMSCs, incubated for 1 day), Group C (PRF+SMSCs, incubated for 3 days), and Group D (PRF+SMSCs, incubated for 5 days). Scanning electron microscopy was used to examine the distribution pattern of SMSCs between groups. The supernatant serum (Group A) and supernatant medium culture (Group D) were collected for the measurement of PDGF, IGF, VEGF, and TGF-β using an enzyme-linked immunosorbent assay sandwich kit. An unpaired t-test was conducted to analyze the differences between Groups A and D (p<0.01). Results: Group D had the most morphologically visible SMSCs attached to the PRF, with elongated and pseudopodia cells. There was a significant difference between the levels of growth factor in Groups A and D (p=0.0001; p<0.01). Conclusion: SMSCs were able to adhere to and distribute evenly on the surface of PRF after 5 days of incubation. The PRF incorporated SMSCs contained high levels of PDGF, IGF, VEGF, and TGF- β, which may prove to have potential in enhancing wound healing