• Title/Summary/Keyword: Collagen matrix gel

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In vitro Angiogenic Activity of Aloe vera Gel on Calf Pulmonary Artery Endothelial (CPAE) Cells

  • Lee, Myoung-Jin;Lee, Ok-Hee;Yoon, Soo-Hong;Lee, Seung-Ki;Chung, Myung-Hee;Park, Young-In;Sung, Chung-Ki;Choi, Jae-Sue;Kim, Kyu-Won
    • Archives of Pharmacal Research
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    • v.21 no.3
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    • pp.260-265
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    • 1998
  • Angiogenic activity of Aloe vera gel was investigated by in vitro assay. We obtained the most active fraction from dichloromethane extract of Aloe vera gel by partitioning between hexane and 90% aqueous methanol. The most active fraction (F3) increased the proliferation of calf pulmonary artery endothelial (CPAE) cells. In addition, F3 fraction induced CPAE cells to invade type I collagen gel and form capillary-like tube through in vitro angiogenesis assay, and increased the invasion of CPAE cells into matrigel through in vitro invasion assay. Furthermore, the effect on the MRNA expression of proteolytic enzymes which are key participants in the regulation of extracellular matrix degradation was investigated by northern blot analysis. F3 fraction enhanced mRNA expression of urokinase-type plasminogen activator (u-PA), matrix metalloproteinase-2 (MMP-2), and membrane-type MMP (MT-MMP) in CPAE cells whereas the expression of plasminogen activator inhibitory (PAl-1) mRNA was not changed.

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Culturing the Human Dental Pulp cells in the Collagen Matrix and on the ground tooth surface (콜라젠 기질(COLLAGEN MATRIX)과 마모된 치아표면에서의 치수세포 배양에 관한 연구)

  • Park, Sang-Hyuk
    • Restorative Dentistry and Endodontics
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    • v.28 no.5
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    • pp.419-424
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    • 2003
  • 이 연구의 목적은 원래의 치수조직과 유사한 조직을 재생하기 위한 pulp tissue engineering의 한 방법으로 건전한 조직으로부터 배양된 치수세포와 쥐의 조섬유세포(NIH 3T3 cell)를 Rat tail type I collagen solution에서 3차원적으로 관찰하기 위한 것으로, 콜라젠 젤의 수축량과 세포의 증식 량을 비교하였으며, 또한 마모된 사람치아의 표면과 배양용기에서 두 세포의 증식 량을 비교하여 다음과 같은 결과를 얻었다. 1. 콜라젠 젤에 NIH 3T3 세포를 배양한 경우 그 수축량은 최소였으나, 치수세포를 배양한 경우 그 수축량은 현저하였다. 2. 서로 다른 수의 치수세포를 콜라젠 젤에서 배양시킨 경우 세포 수가 많을수록 수축량이 증가하였으며, 세포가 없는 콜라젠 젤은 수축하지 않았다. 3. 치수세포를 콜라젠 젤에서 18일간 배양시킨 후 세포의 증식은 거의 없는 반면, NIH 3T3 세포는 계속 증식하였다. 4. 마모된 사람 치아 표면과 배양 용기에서 치수세포와 NIH 3T3세포를 배양한 경우 NIH 3T3세포가 치수세포에 비해 빠르게 증식 하였으며 , 특히 사람 치아의 표면에서 NIH 3T3세포가 현저히 빠른 증식을 보였다. 이상의 결과는 치수세포를 type I collagen gel에서 3차원 적으로 배양 후 치수조직의 재생을 유도하는 pulp tissue engineering에 관한 연구에 발판이 될 것으로 사료된다.

Development of Dermal Equivalent Using Mouse Fibroblasts (세포조직배양법을 이용한 쥐 인공피부의 개발)

  • Yang, Eun-Kyoung;Lee, Jae-Ho;Choe, Tae-Boo;Park, Jung-Keug
    • Microbiology and Biotechnology Letters
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    • v.21 no.4
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    • pp.381-391
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    • 1993
  • As the first stage of development of an artificial skin, fibroblasts were cultured in the collagen matrices to make a living dermal equivalent. Mouse embryonic fibroblasts were incorporated into a collagen matrices on plastic dishes containing concentrated DMEM culture media supplemented with sodium bicarbonate, hepes, antibiotics and fetal bovine serum. As the growth stimulation components, glycosaminoglycans were added: hyaluronic acid, chondroitin sulfate, heparin, chitosan were incorporated into the media at a concentration of either 1% or 5% w/w/ to collagen in order to investigate the effect on development of dermal equivalent. After the few days of incubation, gel matrics were contracted and firm dermal equivalent were formed. And the keratinocytes were cultured on top of dermal equivalent and make a three dimensional artificial skin tissue.

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Tuning the Stiffness of Dermal Fibroblast-encapsulating Collagen Gel by Sequential Cross-linking (연속가교를 통한 피부 진피세포 담지 콜라겐 겔의 강도 제어)

  • Jung, Mun-Hee;Shin, Sung Gyu;Lim, Jun Woo;Han, Sa Ra;Kim, Hee-Jin;Jeong, Jae Hyun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.44 no.1
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    • pp.23-29
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    • 2018
  • In this study, sequential cross-linked collagen gels were successfully prepared with collagen, which is biomaterial, and acrylamide (AAm), which is a synthetic monomer. The elastic moduli (E) of cross-linked collagen gels were increased from 1.5 to 3.0 kPa by varying of AAm concentrations. In addition, human dermal fibroblasts were encapsulated into the porous pores introduced into the gels, and cell growth and behavior were investigated. Increasing E of the gels led to decreases in cell growth rate, while the cellular glycosaminoglycan (GAG) production level was elevated. Overall, the growth and cellular activity of skin cells were influenced by the extracellular matrix properties of the collagen gels. In conclusion, these results will be highly useful for designing reconstructive skins and various tissue engineering researches.

PRODUCTION OF HUMAN PROTEIN TIMP-2: A HIGHLY EFFECTIVE ANTI-AGING INGREDIENT

  • Schutz, R.;Imfeld, D.
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.590-600
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    • 2003
  • The matrix metalloproteinases (MMPs) are a family of enzymes responsible for degrading connective tissue. MMPs catalyze the breakdown of collagen from the extracellular matrix, leading to wrinkle formation and accelerated skin aging. Furthermore, ultraviolet irradiation causes increased expression of certain MMPs. In the extracellular matrix turnover, MMPs are interacting with endogenous regulators named tissue inhibitors of metalloproteinases (TIMPs). Using peptide substrate assays, it has been demonstrated that TIMP-MMP complexes interact highly specifically with $K_{i}$ values of 10$^{-9}$ -10$^{-16}$ M. Therefore applications for TIMP as inhibitor of collagen degradation are suggested for cosmetic anti-aging products to prevent wrinkle formation and loss of elasticity. To date four TIMP proteins (TIMP-1, TIMP-2, TIMP-3 and TIMP-4) have been identified which show a high degree in sequence similarity. The production of human TIMP-2, a 194-residue nonglycosylated protein, was performed by fed-batch culture of Escherichia coli. TIMP-2 accumulated in the bacterial cells in an insoluble form as inclusion bodies. The inclusion bodies were solubilized and the protein refolded to yield the native TIMP-2 in the active form. The integrity of the protein was confirmed by mass analysis, Edman sequencing and gel shift experiments with authentic samples. The inhibitory activity of the refolded and purified TIMP-2 was demonstrated with MMP-1 and MMP-2 assays using synthetic fluorogenic peptide substrates.s.

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ISOLATION OF HUMAN ALVEOLAR BONE-DERIVED CELLS AND IN VITRO AMPLIFICATION FOR TISSUE ENGINEERING (조직공학용 사람 치조골세포의 인공증식)

  • Choi, Byung-Ho;Park, Jin-Hyoung;Huh, Jin-Young;Yoo, Jae-Ha
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.27 no.5
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    • pp.453-456
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    • 2001
  • Background: Autogenous alveolar bone cell transplantation may be suitable for tissue engineering for alveolar bone reconstruction. This study aimed to isolate human alveolar bone-derived cells (HABDCs) and to evaluate the ability of collagen gels to support HABDC proliferation and differentiation for human alveolar bone tissue engineering applications. Method: Cultures of primary HABDCs were established from alveolar bone chips obtained from 10 persons undergoing tooth extraction. These cells were expanded in vitro until passage 3 and used for the in vitro characterization of HABDCs and the in vitro analysis of collagen gels for alveolar bone tissue engineering. Results: Of the 10 attempts made to obtain HABDC cultures, eight were successful. HABDCs expressed the osteoblastic phenotype characterized by alkaline phosphatase activity, osteocalcin expression and the mineralization of the extracellular matrix in vitro. When seeded on collagen gels, HABDCs penetrated into the collagen gel matrices and proliferated inside the gels. Significantly, when HABDCs were embedded into the gels, collagen fibers and mineralization were produced within the gels. Conclusion: This study demonstrates the feasibility of using cultured HABDCs and collagen gels for human alveolar bone tissue engineering applications.

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THE FINITE ELEMENTS ANALYSIS IN THE THREE DIMENSIONAL CELL CULTURE MODEL OF THE COLLAGEN MATRIX ACCORDING TO THE APPLICATION FORCE (교원섬유 기질을 이용한 3차원 세포 배양 모델에 기계적인 힘을 가하는 경우 기질을 통하여 세포에 가해지는 힘의 특성에 대한 비교 분석 연구)

  • Kim, Hyeon-Jong;Rhyu, In-Chul;Park, Jun-Woo;Kim, Seong-Gon
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.34 no.2
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    • pp.151-156
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    • 2008
  • Different kinds of forces can be applied to the biological tissue. The analysis of the applied force is highly important to explain the mechanism of cellular response. In this study, the applied force to the collagen gel was analyzed by the finite elements analysis. The model received two different kinds of static force (compression and tension). The force range was 50g to 400g. In results, von Mises stress was concentrated in the peripheral region in the compression model. It was concentrated in the central area in the tension model. However, the compressive force was high in the peripheral area of the compression model and the tensional force was also high in the same area of the tension model. In conclusion, the applied force could be different to the region and it should be considered in the experiment to analyze the effects of the mechanical force on the cells.

Preparation and Characterization of Alginate-Chitosan Microsphere for Controlled Delivery of Silver Sulfadiazine (설파디아진은의 방출제어를 위한 알지네이트-키토산 미립구의 제조 및 특성)

  • Cho, Ae-Ri
    • Journal of Pharmaceutical Investigation
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    • v.31 no.2
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    • pp.101-106
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    • 2001
  • Alginate-chitosan (anion-cationic polymeric complex) was prepared to control the release rate of silver sulfadiazine (AgSD). Na-alginate (2%) solution containing AgSD was gelled in $CaCl_2$ solution. The gel beads formed were immediately encapsulated with chitosan (CS). The gel matrix and membrane were then reinforced with chondroitin-6-sulfate (Ch6S). Release rate of AgSD from the gel matrix was investigated by placing alginate beads in the sac of cellulose membrane simmered in HEPES-buffer solution. The concentration of AgSD released was analyzed by UV at 264 nm. Incorporation capacity of AgSD in Ca-alginate gel was more than 90%. Alginate-Ch6S-CS could control the release rate of AgSD. The amount of AgSD release was dependent on the AgSD loading dose. Incorporation of tripolyphosphate (polyanionic crosslinker) onto the alginate-Ch6S-CS bead increased the release rate of AgSD. Collagen-coating had no influence on the AgSD release rate. Alginate-Ch6S-CS beads with a sufficiently high AgSD encapsulation were capable of controlling the release of the drug over 10 days. In summary, alginate-Ch6S-CS beads could be used as a sustained delivery for AgSD and provide local targeting with low silver toxicity and patient discomfort.

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The Effects of Fetal Bovine Serum, Epidermal Growth Factor, and Retinoic Acid on Adult Rat Islets Embedded in Collagen Gels

  • Shin, Jun-Seop;Chang, Hyo-Ihl;Sung, Ha-Chin;Kim, Chan-Wha
    • Journal of Microbiology and Biotechnology
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    • v.9 no.2
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    • pp.150-156
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    • 1999
  • The induction of proliferation of adult rat islets was investigated under various culture conditions. The islets were isolated from male Sprague-Dawley rats and subsequently embedded in collagen gels, which mimic the in vivo three-dimensional surroundings. During the culture period, the effects of heterologous serum (fetal bovine serum, FBS), epidermal growth factor (EGF), and retinoic acid (RA) on islet growth were examined with respect to the morphological and total DNA content changes. To investigate these changes at the cellular level, whole mount immunocytochemistry using specific antibodies for insulin and glucagon was performed. The results showed that (i) collagen gels as an extracellular matrix can maintain islets in a similar way to that in vivo, (ii) heterologous serum (FBS) had stimulatory effects on islet proliferation in a dose-dependent manner, (iii) RA had inhibitory effects on islet proliferation induced by the serum in a dose-dependent manner, (iv) EGF had weak inhibitory effects on islet proliferation induced by the serum except at the concentration of 10 nM where its effect was not significant, and (v) whole mount immunocytochemistry revealed that newly proliferated islet cells were mainly $\beta$-and $\alpha$-cells.

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