In Vitro Characterization of Poly-Glycolyc Lactic-Co Acid (PLGA) –Collagen Based on Red Snapper Fish Scales (Lutjanus Sp.) Coating Chitosan as Duramater Artificial Candidate

Head trauma was the third cause of deaths that have a high rank that can make serious head injury for 25.5%-54.9%. This study has been conducted by making a replacement layer of the brain (dura) to overcome the impact of dural defect by utilizing waste fish scales red snapper (Lutjanus sp.). Synthesis brain membranes lining processed by casting method with each various concentrations of chitosan coating of 1%, 1.5%, and 2% then dried using vacuum dry. The samples then were characterized by tensile test, FTIR, SEM and MTT Assay. FTIR test results showed that red snipperscales can produce collagen powder at amide A group with stretching of –NH functional group, amide B group has stretching of CH2 assymetry, amide I area, amide II and amide III area which show –NH bonding. Tensile test results showed that the combination between PLGA-Collagen Chitosan Coating 2% produced the highest tensile strength is 4.8 MPa which meet the standards of human duramater strength. MTT Assay results showed that the dural membrane produced no toxic seen from living cells reached 98.32%. Poly - Glycolyc Lactic - Co Acid (PLGA) - collagen coating chitosan based on red snapper fish scales (Lutjanus sp.) composites has potency as duramater artificial candidate due to the chemistry, biological and physical characteristics.

In Vitro Characterization of Poly-Glycolyc Lactic-Co Acid (PLGA) –Collagen Based on Red Snapper Fish Scales (Lutjanus Sp.) Coating Chitosan as Duramater Artificial Candidate

Head trauma is the third cause of deaths with a high rank, causing severe head injury for 25.5%-54.9%. This study has been conducted by making a replacement layer of the brain (dura) to overcome the dural defect's impact by utilizing waste fish scales red snapper (Lutjanus Sp.). Synthesis brain membranes lining processed by casting method with each various chitosan coating concentrations of 1%, 1.5%, and 2%, then dried using vacuum dry. The samples were then characterized by tensile tests, FTIR, SEM, and MTT Assay. FTIR test results showed that red snipper scales could produce collagen powder at amide A group with stretching of –NH functional group, amide B group, has stretching of CH2 asymmetry, amide I area, amide II, and amide III area, which show –NH bonding. Tensile test results showed that the combination of PLGA-Collagen Chitosan Coating 2% produced the highest tensile strength is 4.8 MPa, which met human dura mater strength standards. MTT Assay results showed that the dural membrane produced no toxic seen from living cells reached 98.32%. Poly - Glycolic Lactic - Co Acid (PLGA) - collagen coating chitosan-based on red snapper fish scales (Lutjanus Sp.) composites has potency dura mater, artificial candidate, due to the chemistry, biological, and physical characteristics.

The effects of coating culture dishes with collagen on fibroblast cell shape and swirling pattern formation

Motile human-skin fibroblasts form macroscopic swirling patterns when grown to confluence on a culture dish. In this paper, we investigate the effect of coating the culture-dish surface with collagen on the resulting pattern, using human-skin fibroblast NB1RGB cells as the model system. The presence of the collagen coating is expected to enhance the adherence of the fibroblasts to the dish surface, and thereby also enhance the traction that the fibroblasts have as they move. We find that, contrary to our initial expectation, the coating does not significantly affect the motility of the fibroblasts. Their eventual number density at confluence is also unaffected. However, the coherence length of cell orientation in the swirling pattern is diminished. We also find that the fibroblasts cultured in collagen-coated dishes are rounder in shape and shorter in perimeter, compared with those cultured in uncoated polystyrene or glass culture dishes. We hypothesise that the rounder cell-shape which weakens the cell–cell nematic contact interaction is responsible for the change in coherence length. A simple mathematical model of the migrating fibroblasts is constructed, which demonstrates that constant motility with weaker nematic interaction strength does indeed lead to the shortening of the coherence length.

Frame Coating of Single-Walled Carbon Nanotubes in Collagen on PET Fibers for Artificial Joint Ligaments

The coating formation technique for artificial knee ligaments was proposed, which provided tight fixation of ligaments of polyethylene terephthalate (PET) fibers as a result of the healing of the bone channel in the short-term period after implantation. The coating is a frame structure of single-walled carbon nanotubes (SWCNT) in a collagen matrix, which is formed by layer-by-layer solidification of an aqueous dispersion of SWCNT with collagen during spin coating and controlled irradiation with IR radiation. Quantum mechanical method SCC DFTB, with a self-consistent charge, was used. It is based on the density functional theory and the tight-binding approximation. The method established the optimal temperature and time for the formation of the equilibrium configurations of the SWCNT/collagen type II complexes to ensure maximum binding energies between the nanotube and the collagen. The highest binding energies were observed in complexes with SWCNT nanometer diameter in comparison with subnanometer SWCNT. The coating had a porous structure—pore size was 0.5—6 μm. The process of reducing the mass and volume of the coating with the initial biodegradation of collagen after contact with blood plasma was demonstrated. This is proved by exceeding the intensity of the SWCNT peaks G and D after contact with the blood serum in the Raman spectrum and by decreasing the intensity of the main collagen bands in the SWCNT/collagen complex frame coating. The number of pores and their size increased to 20 μm. The modification of the PET tape with the SWCNT/collagen coating allowed to increase its hydrophilicity by 1.7 times compared to the original PET fibers and by 1.3 times compared to the collagen coating. A reduced hemolysis level of the PET tape coated with SWCNT/collagen was achieved. The SWCNT/collagen coating provided 2.2 times less hemolysis than an uncoated PET implant. MicroCT showed the effective formation of new bone and dense connective tissue around the implant. A decrease in channel diameter from 2.5 to 1.7 mm was detected at three and, especially, six months after implantation of a PET tape with SWCNT/collagen coating. MicroCT allowed us to identify areas for histological sections, which demonstrated the favorable interaction of the PET tape with the surrounding tissues. In the case of using the PET tape coated with SWCNT/collagen, more active growth of connective tissue with mature collagen fibers in the area of implantation was observed than in the case of only collagen coating. The stimulating effect of SWCNT/collagen on the formation of bone trabeculae around and inside the PET tape was evident in three and six months after implantation. Thus, a PET tape with SWCNT/collagen coating has osteoconductivity as well as a high level of hydrophilicity and hemocompatibility.

PC12 Cell Line: Cell Types, Coating of Culture Vessels, Differentiation and Other Culture Conditions

The PC12 cell line is one of the most commonly used in neuroscience research, including studies on neurotoxicity, neuroprotection, neurosecretion, neuroinflammation, and synaptogenesis. Two types of this line are available in the ATCC collection: traditional PC12 cells grown in suspension and well-attached adherent phenotype. PC12 cells grown in suspension tend to aggregate and adhere poorly to non-coated surfaces. Therefore, it is necessary to modify the surface of culture vessels. This paper aims to characterise the use of two distinct variants of PC12 cells as well as describe their differentiation and neuronal outgrowth with diverse NGF concentrations (rat or human origin) on various surfaces. In our study, we evaluated cell morphology, neurite length, density and outgrowth (measured spectrofluorimetrically), and expression of neuronal biomarkers (doublecortin and NeuN). We found that the collagen coating was the most versatile method of surface modification for both cell lines. For adherent cells, the coating was definitely less important, and the poly-d-lysine surface was as good as collagen. We also demonstrated that the concentration of NGF is of great importance for the degree of differentiation of cells. For suspension cells, we achieved the best neuronal characteristics (length and density of neurites) after 14 days of incubation with 100 ng/mL NGF (change every 48 h), while for adherent cells after 3–5 days, after which they began to proliferate. In the PC12 cell line, doublecortin (DCX) expression in the cytoplasm and NeuN in the cell nucleus were found. In turn, in the PC12 Adh line, DCX was not expressed, and NeuN expression was located in the entire cell (both in the nucleus and cytoplasm). Only the traditional PC12 line grown in suspension after differentiation with NGF should be used for neurobiological studies, especially until the role of the NeuN protein, whose expression has also been noted in the cytoplasm of adherent cells, is well understood.

Collagen Coating for Tissue Culture

This protocol is intended for collagen coating of tissue culture dishes and Transwell® multiple well plates. The attached methods document is a formal version of the information included here.Disclaimer: The information presented here has been reviewed and approved for publication by the US Environ do not necessarily represent Agency policy. Mention of trade names or commercial products does not constitute endorsement or recommendations for use.