Bio-Fabrication of Human Amniotic Membrane Zinc Oxide Nanoparticles and the Wet/Dry HAM Dressing Membrane for Wound Healing

Publication history: Accepted - 25 June 2021; Published online - 28 July 2021.The preparation of unique wet and dry wound dressing products derived from unprocessed human amniotic membrane (UP-HAM) is described. The UP-HAM was decellularized, and the constituent proteins were cross-linked and stabilized before being trimmed and packed in sterile Nucril-coated laminated aluminium foil pouches with isopropyl alcohol to manufacture processed wet human amniotic membrane (PWHAM). The dry type of PD-HAM was prepared by decellularizing the membrane, UV irradiating it, lyophilizing/freeze-drying it, sterilizing it, and storing it at room temperature. The UP-HAM consists of a translucent yellowish mass of flexible membranes with an average thickness of 42 µm. PW-HAM wound dressings that had been processed, decellularized, and dehydrated had a thinner average thickness of 30 µm and lacked nuclear-cellular structures. Following successful decellularization, discrete bundle of fibrous components in the stromal spongy layers, microvilli and reticular ridges were still evident on the surface of the processed HAM, possibly representing the location of the cells that had been removed by the decellularization process. Both wet and dry HAM wound dressings are durable, portable, have a shelf life of 3–5 years, and are available all year. A slice of HAM dressing costs 1.0 US/cm2 . Automation and large-scale HAM membrane preparation, as well as storage and transportation of the dressings, can all help to establish advanced technologies, improve the efficiency of membrane production, and reduce costs. Successful treatment of wounds to the cornea of the eye was achieved with the application of the HAM wound dressings. The HAM protein analysis revealed 360 µg proteins per gram of tissue, divided into three main fractions with MWs of 100 kDa, 70 kDa, and 14 kDa, as well as seven minor proteins, with the 14 kDa protein displaying antibacterial properties against human pathogenic bacteria. Frontiers in Bioengineering and Biotechnology | www.frontiersin.org 1 July 2021 | Volume 9 | Article 695710 fbioe-09-695710 July 22, 2021 Time: 16:39 # 2 Ramasamy et al. HAMP-ZnO Nanoparticles HAM Wound Dressing Wet and dry wound dressings were produced. HAM proteins were purified and analysed. The zinc oxide nanoparticles (HAMP-ZnO NP) made from HAM proteins were characterised and tested for their antibacterial activity. Wounds to the cornea of the eye healed easily when treated with HAM wound dressings. Fresh human Amniotic membrane, Serological screening, selection of disease-free HAM, reome stromal layer, preparation of HAM. UNPROCESSED HAM Cuboidal epithelial cells, basement membrane, compact layer, stromal and spongy layers containing scatted fibroblast cells are visible in hsitological analysis. The flow chart depicts the methods for processing, and preparation of wet (PWHAM) and dry (PD-HAM) wound healing dressings. HAM proteins, Nanoparticle synthesis (HAMP-ZnO NP) and analysis. Antibacterial analysis show Inhibition of growth and biofilm formation of pathogenic bacteria . Processed HAM lacked a nuclear-cellular epithelium, but it did have a distinct fibrous elements in basement membrane, stromal and spongy layers. Processed PW-HAM (Light &SEM) showed smooth epithelial surface topography with microvilli,. HAM dressing, wet/dry, packed, labelled, sterilised and processed. They are durable, portable, have long shelf life . A slice of HAM dressing costs US 1.0 / cm² . The wound dressings are ready to be applied. The dermal wounds and conjunctival surface can be successfully repaired using processed HAM wound dressings GRAPHICAL ABSTRACT | Flow chart depicting the methods, preparing, and characterizing, by histological, and scanning electron microscopy, of wet (PW-HAM) and dry (PD-HAM)of wound healing dressing, and preparation of nanoparticles (HAMP ZnO NP); and application of HAM wound dressing. A wide range of antibacterial activity was observed after treatment with 75 µg/ml zinc oxide nanoparticles derived from human amniotic membrane proteins (HAMP-ZnO NP), including dose-dependent biofilm inhibition and inhibition of Gram-positive (S. aureus, S. mutans, E. faecalis, and L. fusiformis) and Gram-negative bacteria (S. sonnei, P. aeruginosa, P. vulgaris, and C. freundii).PR has acknowledged Sree Balaji Medical College and Hospital for providing the article processing charges of the journal, and moral and technical support. The support of Cologenesis Health Care Pvt. Ltd. for a study on “Human amniotic membrane for ocular and dermal applications” is sincerely appreciated

Open Access Article│www.njcmindia.org pISSN 0976 3325│eISSN 2229 6816 National Journal of Community Medicine│Volume 4│Issue 3│July -Sept 2013 Kancheepuram District of Tamil Nadu, India and their Association with Risk Factors of Cardiovascular Diseases

ABSTRACT Background: The proportion of hypertension in India as reported by various literatures has been on an increasing trend for the last three decades. The present study was carried out to determine the proportion of hypertension, its associated risk factors as well as to increase the awareness on importance of life style modifications among people visiting a rural hospital

Thyroid-Stimulating Hormone Favors Runx2-Mediated Matrix Mineralization in HOS and SaOS2 Cells: An In Vitro and In Silico Approach

Osteoporosis is a skeletal disease that is both systemic and silent characterized by an unbalanced activity of bone remodeling leading to bone loss. Rising evidences demonstrate that thyroid stimulating hormone (TSH) has an important role in the regulation on the metabolism of bone. However, TSH regulation on human osteoblast essential transcriptional factors has not been identified. Current study examined the role of TSH on human osteoblastic Runx2 expression and their functional genes by in vitro and in slico analysis. Human osteoblast like (HOS and SaoS-2) cells were cultured with DMEM and treated with hTSH at the concentration of 0.01 ng/mL and 10 ng/mL. After treatment, osteoblastic Runx2 and IGF-1R beta expression were studied using RT-PCR and western blot analysis. TSH treatment induced osteoblastic essential transcriptional factor, Runx2 in HOS and SaOS2 cells on 48 h duration and elevated the expression of IGF-IR β gene and Protein in SaoS-2 cells. TSH also promotes Runx2 responsive genes such as ALP, Collagen and osteocalcin in SaOS2 cells on day 2 to day 14 of 10 ng/mL of treatment and favors’ matrix mineralization matrix in these cells. In addition, TSH facilitated human osteoblastic cells to mineralize their matrix confirmed by day 21 of alizarin red calcium staining. In silico study was performed to check CREB and ELK1 interaction with Runx2. Results of in silico analysis showed that TSH mediated signalling molecules such as CREB and ELK1 showed interaction with Runx2 which involve in osteobalstic gene expression and differentiation. Present findings confirm that TSH promotes Runx2 expression, osteoblastic responsive genes and bone matrix formation