Correlation of spontaneous adipocyte generation with osteogenic differentiation of porcine skin-derived stem cells

The objective of this study was to examine effects of spontaneous adipocyte generation on osteogenic differentiation of porcine skin-derived stem cells (pSSCs). Correlation between osteogenic differentiation and adipocyte differentiation induced by osteocyte induction culture was determined using different cell lines. Osteogenic differentiation efficiency of pSSCs was then analyzed by controlling the expression of adipocyte-specific transcription factors during osteogenic induction culture. Among four cell lines, pSSCs-II had the lowest lipid droplet level but the highest calcium content (p < 0.05). It also expressed significantly low levels of peroxisome proliferator-activated receptor gamma 2 (PPARγ2) and adipocyte protein 2 (aP2) mRNAs but very high levels of runt-related transcription factor 2 (Runx2) and alkaline phosphatase (ALP) mRNAs as osteogenic makers (p < 0.05). Oil red O extraction was increased by 0.1 µM troglitazone (TGZ) treatment but decreased by 50 µM bisphenol A diglycidyl ether (BADGE) (p < 0.05). Calcium content was drastically increased after BADGE treatment compared to that in osteogenic induction control and TGZ-treated pSSCs (p < 0.05). Relative expression levels of PPARγ2 and aP2 mRNAs were increased by TGZ but decreased by BADGE. Expression levels of Rucx2 and ALP mRNAs, osteoblast-specific marker genes, were significantly increased by BADGE treatment (p < 0.05). The expression level of BCL2 like 1 was significantly higher in BADGE-treated pSSCs than that in TGZ-treated ones (p < 0.05). The results demonstrate that spontaneous adipocyte generation does not adversely affect osteogenic differentiation. However, reducing spontaneous adipocyte generation by inhibiting PPARγ2 mRNA expression can enhance in vitro osteogenic differentiation of pSSCs.

Revealing Weapon Impacts on Clothes Using Reaction Reagents for Amino Acids / 대한법의학회지

If we analogize any external physical force applied to victims of crimes involving violence, it would be possible to not only presume the mutual action between victims and suspects but also deduce more facts related to the cases. Therefore, in this study, defining the phenomenon of amino acid compounds in sweat spreading into clothes as impact marks, experiments using ninhydrin, 1,8-dizafluoren-9-one (DFO), 1,2-indanedione-zinc (1,2-IND-Zn) were conducted to determine developmental variations through change over time, which was not performed in previous studies. A 5-week period was set up including first damage as a variation factor, and materials in each action were developed using certain reagents. The level of specimen development depending on the change over time was identified. Thus, no changes were observed at each initial level of development.

An experimental study of bio-polymer composite film as a micro-skin autograft covering / 实用医学杂志

Objective To explore the promoting effects of bio-polymer composite film as a micro-skin auto-graft covering on wound healing. Methods The full thickness skin defect models were made on both sides of 30 experimental rabbits. Then, the rabbits were randomly divided into experimental group and control group. In the for-mer group, the side was covered with chitosan/glucomannan composite membrane and in the latter, the side cov-ered with acelluar porcine skin after micro-skin autograft. We obtained wound tissues at week 1 , 2, 3, 4 and 5 af-ter operation. The conditions of wound healing were observed, the rate of wound healing was calculated, HE stain-ing was made, and PCNA and CD31 were detected by immunohistochemistry. Results (1) During 2~4 weeks af-ter operation, the rate of wound healing in the experimental group was significantly higher than that of the control wound (P<0.01). (2) The amount of neutrophil in experimental group was less than that of the control after oper-ation. (3) During 1 ~ 2 weeks after operation, the expression of PCNA in the experimental group was higher than that of the control group (P < 0.01), but lower than the control wounds during 1 ~ 2 weeks after operation (P <0.01). (4) During 1 ~ 5 weeks after operation, the expression of CD31 in the experimental group was higher than that of the control group (P < 0.01). Conclusion Chitosan/glucan-mannan composite membrane as a micro-skin autograft covering may promote wound healing.

A cellular porcine skin and allogeneic skin as wound-covering materials for extensive deep burns: A comparative study / 第二军医大学学报

Objective: To evaluate the advantages of microskin graft using acellular porcine skin for treatment of extensive deep burns by comparing with that using allogeneic skin. Methods: A retrospective analysis was conducted on 70 severe burn patients who were treated in the Second Affiliated Hospital of Kunming Medical University during Jan. 1999 to Jan. 2011. The patients were divided into the acellular porcine skin group and allogeneic skin group, each containing 35 patients. The survival rates of microskin grafts were determined at 4 weeks post-operation. Besides, the rejection of acellular porcine skin and allogeneic skin, changes of body temperature, white blood cell (WBC) count, lymphocyte and serum protein were observed at pre- and post-operation. Results: (1) The survival rate was (71. 5 ± 6. 6)% in acellular porcine skin group and (70. 6 ± 7. 5)% in allogeneic skin group, with no significant difference found between the two groups (P>0. 05). (2) Acellular porcine skin group. At 3 days post-operation the acellular porcine skin was still attached to the wound, most of the skin was not discolorated, and small part of the skin became cinnamomeous. The acellular porcine skin was gradually separated from the auto-microskin at 3-4 weeks post operation, and there was small amount of exudates under the acellular porcine skin, which could be drained through a small cut. In the pressed area, there was still a small amount of exudates, but the acellular porcine skin was not dissolved and the microskin grafts survived and became confluent. (3) Allogeneic skin group. The allogeneic epidermal was rejected and was off from the wound at 3-14 days post transplantation, and at 10-30 days after transplantation the allogenic dermis became dry. During 25-60 days after transplantation, the allogenic dermis was completely stripped off, the microskin grafts became confluent, and the wound was healed. (4) The body temperature of the two groups was significantly descended after operation (P0. 05). Conclusion: Microskin graft using acellular porcine skin, instead of allogeneic skin, for extensive burn patients can inhibit systematic inflammatory response, improve the nutrition condition, and reduce the using of allogeneic skin. Acellular porcine skin might be a suitable alternative for allogeneic skin.

Fluorometric quantification of protoporphyrin IX in biological skin samples from in vitro penetration/permeation studies

A fluorometric analytical method was developed for quantification of protoporphyrin IX (PpIX) in skin samples and receptor phase solution after in vitro cutaneous penetration/permeation studies. Analytical conditions used were: excitation and emission wavelengths: 400 nm and 632 nm; bandwidth: 0.5 nm; excitation and emission slits: 10/10. PpIX was recovered from two different layers of skin, the stratum corneum (SC) and the epidermis plus dermis ([E+D]), by vortex homogenization, probe and bath sonication, using DMSO as an extraction solvent. The detection and quantification limits were 0.002 and 0.005 μg/mL, respectively. The assay was linear from 0.005 - 0.5 μg/mL. The within-day and between-day assay precision and accuracy in DMSO and receptor phase solution were each studied at the two concentration levels 0.04 and 0.2 μg/mL, and 0.01 and 0.08 μg/mL, respectively. The coefficients of variation and deviation from the theoretical values were lower than 5%. The skin recovery of PpIX from SC and [E+D] layers using two different concentrations (0.5 and 1.0 μg/mL) were all above 90.0%. The method described has potential application to in vitro penetration/permeation studies of PpIX using porcine skin as a biological membrane model.

Um método analítico por espectrofluorimetria foi desenvolvido para quantificar a protoporfirina IX (Pp IX) em amostras de pele e fase receptora após a realização de testes in vitro de penetração/permeação cutâneas. As condições analíticas utilizadas foram: comprimentos de onda de excitação e emissão: 400 nm e 632 nm; largura de banda: 0,5 nm; fendas de excitação e emissão: 10/10. A PpIX foi extraída de amostras de estrato córneo (EC) e da epiderme sem estrato córneo + derme ([E+D]) através da agitação em vórtex e sonicação por haste e banho, utilizando-se o DMSO como solvente extrator. O limite de detecção e quantificação foram, respectivamente, de 0,002 e 0,005 μg/mL. O método mostrou-se linear da faixa de 0,005 - 0,5 μg/mL. A precisão e exatidão intra e inter-ensaio em DMSO e na fase receptora foram validadas utilizando-se duas concentrações distintas, respectivamente, de 0,004 e 0,2 μg/mL, e 0,01 e 0,08 μg/mL. Os valores de coeficiente de variação e o desvio do valor teórico foram inferiores a 5%. A recuperação da PpIX das camadas da pele (EC e [E+D]) utilizando-se duas concentrações distintas (0,5 e 1,0 μg/mL) foram todas acima de 90,0%. O método descrito pode ser utilizado para determinação da PpIX após estudos de penetração/permeação cutânea in vitro utilizando pele de porco como modelo de membrana.