Effect of Stromal Vascular Fractions on Angiogenesis of Injected Diced Cartilage.

OBJECTIVES: This study explored the effect of adipose-derived stromal vascular fractions (SVFs) on angiogenesis in injected autologous diced cartilage. METHODS: Stromal vascular fractions were extracted by enzymatic digestion. Cartilage grafts were harvested from 1 side of the auricular cartilage of New Zealand rabbit and then diced to a size of 1.0 mm3. The grafts were divided into 2 groups. The control group was diced cartilage mixed with culture medium, and the experimental group was diced cartilage mixed with SVFs. The 2 groups of composite grafts were subcutaneously implanted on both sides of the back of each rabbit. After 4, 12 and 24 weeks, the tissue structure, number of blood vessels, and angiogenic factors in the grafts were observed. RESULTS: The SVFs conformed to the current standard of the biological evaluation. Under an inverted microscope, the number of layers of chondrocytes in the experimental group was higher than that in the control group at 4 weeks. A small number of inflammatory cells and blood vessels were observed around the cartilage grafts. At 12 and 24 weeks, the volume of tissue was increased gradually by general observation. And a large number of chondrocytes were observed microscopically, whereas the number of inflammatory cells decreased. And meanwhile additional new blood vessels were observed. Immunohistochemical analysis of CD31 showed that the number of capillaries in the control group was significantly lower than that in the experimental group at 4, 12 and 24 weeks. Further, the expression of Hypoxia inducible factor-1 (HIF-1) and vascular endothelial growth factor (VEGF) mRNA and protein were measured by RT-PCR and Western bloting, respectively. The results showed that the mRNA expression of VEGF and HIF-1α in the experimental group was increased. The mRNA level remained higher than that of the control group at 24 weeks (P < 0.05). And the relative expression levels of VEGF and HIF-1α protein in the experimental group were higher than those in the control group at 4, 12 and 24 weeks (P < 0.05). CONCLUSION: Autologous diced cartilage mixed with adipose-derived SVFs can promote angiogenesis when transplanted by injection. Further research showed that SVFs could increase the expression levels of VEGF and HIF-1α in the grafts, which may be part of the mechanism that SVFs promoted the angiogenesis of diced cartilage.

Identification of transcriptomic characteristics during nasal capsular contracture progression using RNA deep sequencing.

Nasal capsular contracture is a prevalent complication commonly observed after rhinoplasty. However, the mechanism underlying the pathogenesis of nasal capsular contracture is largely unclear compared to that of breast capsular contracture. This study aimed to identify the key genes implicated in nasal capsular contracture progression using RNA deep sequencing (RNA-seq). Biopsy samples were taken from Grade II to Grade IV nasal fibrous capsular tissues. The former is regarded as the relatively normal tissues and thus was set as control group, while the latter was treated as pathological group. Results from RNA-seq underwent GO enrichment and KEGG pathway analysis and subsequent verification by quantitative reverse transcriptase polymerase chain reaction and western blot assays. RNA-seq analysis showed that 3149 genes were up-regulated and 3131 genes in pathological groups compared with controls. The top 30 up-regulated genes included many chemokines (e.g., CCL18, CCL13, CCL17 and CCL8), matrix metallopeptidases (e.g., MMP9 and MMP12) and integrin proteins (e.g., ITGAM and ITGB2). GO enrichment analysis demonstrated that the up-regulated genes affected various immune functions, including immune system process, cell activation, leukocyte activation, defence response and positive regulation of immune. The down-regulated gene primary influenced muscle development and functions as well as metabolic processes. In summary, this study reveal that abnormal changes of immune functions, muscle develop and metabolic processes are probably implicated in the pathogenesis of nasal capsular contracture.

Hyperactivation of RAP1 and JAK/STAT Signaling Pathways Contributes to Fibrosis during the Formation of Nasal Capsular Contraction.

Silicone implant-based augmentation rhinoplasty or mammoplasty induces capsular contracture, which has been acknowledged as a process that develops an abnormal fibrotic capsule associated with the immune response to allogeneic materials. However, the signaling pathways leading to the nasal fibrosis remain poorly investigated. We aimed to explore the molecular mechanism underlying the pathogenesis of nasal capsular contracture, with a specific research interest in the signaling pathways involved in fibrotic development at the advanced stage of contracture. By examining our recently obtained RNA sequencing data and global gene expression profiling between grade II and grade IV nasal capsular tissues, we found that both the RAP1 and JAK/STAT signaling pathways were hyperactive in the contracted capsules. This was verified on quantitative real-time PCR which demonstrated upregulation of most of the representative component signatures in these pathways. Loss-of-function assays through siRNA-mediated Rap1 silencing and/or small molecule-directed inhibition of JAK/STAT pathway in ex vivo primary nasal fibroblasts caused a series of dramatic behavioral and functional changes, including decreased cell viability, increased apoptosis, reduced secretion of proinflammatory cytokines, and synthesis of type I collagen, compared to control cells, and indicating the essential role of the RAP1 and JAK/STAT signaling pathways in nasal capsular fibrosis. Our results sheds light on targeting downstream signaling pathways for the prevention and therapy of silicone implant-induced nasal capsular contracture.

Viability and Biomechanics of Bare Diced Cartilage Grafts in Experimental Study.

OBJECTIVE: To assess the viability and biomechanics of bare diced cartilage grafts. METHODS: Cartilage samples were collected from 1 ear in 15 rabbits as well as costal cartilage. Each rabbit was inserted bare diced- and single-strip costal-cartilage grafts, respectively, into paraspinal subcutaneous pockets: after euthanasia at 2 months, specimens were weighed, with diced cartilage grafts examined histomorphologically by hematoxylin-eosin staining, masson trichrome staining, and immunohistochemistry. Finally, biomechanical properties of grafts were assessed. RESULTS: Bare diced cartilage grafts were connected into an integrated mass after 2 months, and inward growth of fibrous tissues and angiogenesis were observed. Mean wet weights of diced cartilage grafts were 1.603 ±â€Š0.278 and 1.662 ±â€Š0.204 g pre- and postoperation, respectively; those of costal cartilage grafts were 0.053 ±â€Š0.008 and 0.058 ±â€Š0.008 g, respectively. In compression assays, mean modulus values of elasticity at yield in diced- and costal-cartilage grafts were 7.65 ±â€Š0.59 and 22.30 ±â€Š1.15 MPa, respectively (P < 0.05); mean stress values were 4.07 ±â€Š0.38 and 12.50 ±â€Š1.15 MPa, respectively (P < 0.05). In the tensile test, mean modulus values of elasticity at yield of diced- and costal-cartilage grafts were 4.70 ±â€Š0.78 and 10.59 ±â€Š1.39 MPa, respectively (P < 0.05), mean stress values were 0.82 ±â€Š0.05 and 1.76 ±â€Š0.21 MPa, respectively (P < 0.05). CONCLUSIONS: Diced cartilage grafts had favorable viability and growth. Despite reduced elasticity and stress values, they still can be served as substitute for supportive filling materials.

Viability and Biomechanics of Diced Cartilage Blended With Platelet-Rich Plasma and Wrapped With Poly (Lactic-Co-Glycolic) Acid Membrane.

OBJECTIVES: The objective of this study was to investigate the viability and biomechanics of diced cartilage blended with platelet-rich plasma (PRP) and wrapped with poly (lactic-co-glycolic) acid (PLGA) membrane in a rabbit model. METHODS: A total of 10 New Zealand rabbits were used for the study. Cartilage grafts were harvested from 1 side ear. The grafts were divided into 3 groups for comparison: bare diced cartilage, diced cartilage wrapped with PLGA membrane, and diced cartilage blended with PRP and wrapped with PLGA membrane. Platelet-rich plasma was prepared using 8 mL of auricular blood. Three subcutaneous pockets were made in the backs of the rabbits, and the grafts were placed in these pockets. The subcutaneous implant tests were conducted for safety assessment of the PLGA membrane in vivo. All of the rabbits were sacrificed at the end of 3 months, and the specimens were collected. The sections were stained with hematoxylin and eosin, toluidin blue, and collagen II immunohistochemical. Simultaneously, biomechanical properties of grafts were assessed. RESULTS: This sample of PLGA membrane was conformed to the current standard of biological evaluation of medical devices. Moderate resorption was seen at the end of 3 months in the gross assessment in diced cartilage wrapped with PLGA membrane, while diced cartilage blended with PRP had no apparent resorption macroscopically and favorable viability in vivo after 3 months, and the histological parameters supported this. Stress-strain curves for the compression test indicated that the modulus of elasticity of bare diced cartilage was 7.65 ±â€Š0.59 MPa; diced cartilage wrapped with PLGA membrane was 5.98 ±â€Š0.45 MPa; and diced cartilage blended with PRP and wrapped with PLGA membrane was 7.48 ±â€Š0.55 MPa, respectively. CONCLUSIONS: Diced cartilage wrapped with PLGA membrane had moderate resorption macroscopically after 3 months. However, blending with PRP has beneficial effects in improving the viability of diced cartilages. Additionally, the compression modulus of diced cartilage blended with PRP and wrapped with PLGA membrane was similar to bare diced cartilage.

Induction of melasma by 1064-nm Q-switched neodymium:yttrium-aluminum-garnet laser therapy for acquired bilateral nevus of Ota-like macules (Hori nevus): A study on related factors in the Chinese population.

Laser treatment has emerged as a common treatment modality for acquired bilateral nevus of Ota-like macules (ABNOM). To identify the ratio of melasma induction and exacerbation before and after laser therapy for ABNOM and to observe the risk factors related to the induction and exacerbation of melasma by laser therapy, we analyzed related factors of 1268 adult Chinese patients who underwent 1064-nm Q-switched neodymium:yttrium-aluminum-garnet (Nd:YAG) laser (QNYL) treatment using case series and case-control studies. Overall, 24.0% of the ABNOM patients had mixed melasma. Among the ABNOM patients without melasma, after laser therapy the development of melasma was more frequently noted in patients older than 35 years (P < 0.0001), as well in patients whose ABNOM was less than 10 cm(2) (P = 0.027), ABNOM were light (similar to yellow-brown) in color (P = 0.021) and skin types were closer to type IV (P < 0.0001). New melasma lesions also appeared most frequently in the zygomatic region (P < 0.0001). Among the ABNOM patients with melasma, 89.5% experienced worsening of their melasma, irrespective of their related factors above. We concluded that the risk of inducing melasma is great after 1064-nm QNYL treatment in ABNOM patients, and particularly in the patients with both ABNOM and melasma. ABNOM patients should be treated as early as possible and before the age of 35 years.

17beta-estradiol potentiates ischemia-reperfusion injury in diabetic ovariectomized female rats.

UNLABELLED: To investigate the effect of 17beta-estradiol (E2) on ischemia-reperfusion (I/R) injury in diabetic ovariectomized female rats. Streptozotocin(STZ)-induced diabetic female rats received E2 treatment for 2 weeks after ovariectomy (OVX). A period of 90 min of temporary middle cerebral artery occlusion (tMCAO) was used for the study. Rats were evaluated for physiological data including plasma glucose, E2, MAP, PaCO2 and PaO2 before and after tMCAO. P-selectin expression, myeloperoxidase (MPO) enzyme activity and the cerebral infarct volume were analyzed. RESULTS: The infarct volume in the E2-treated OVX rats is bigger than that in intact and OVX groups. However, there is not a significant different area of cerebral infarct between diabetic OVX and intact rats. Significant upregulation of P-selectin expression and MPO activity of the ischemia-reperfusion hemisphere were observed in E2 + OVX, intact and OVX groups at 8, 24, 72 h in time manner after tMCAO compared with that of the contralateral hemisphere of cerebral ischemia-reperfusion. Both P-selectin expression and MPO activity in the E2 + OVX and intact rats are significantly higher than that in the untreated OVX rats. Chronic estrogen replacement therapy (ERT) potentiates the I/R injury in diabetes female rats. This may be related to the increased expression of P-selectin and MPO activity.