Objective To investigate the effects of the misshapen auricular chondrocytes from microtia in inducing chondrogenesis of human adipose derived stem cells (ADSCs) in vitro. Methods Human ADSCs at passage 3 and misshapen auricular chondrocytes at passage 2 were harvested and mixed at a ratio of 7 ∶ 3 as experimental group (group A, 1.0 × 106 mixed cells). Misshapen auricular chondrocytes or ADSCs at the same cell number served as control groups (groups B and C, respectively). All samples were incubated in the centrifuge tubes. At 28 days after incubation, the morphological examination was done and the wet weight was measured; the content of glycosaminoglycan (GAG) was detected by Alcian blue colorimetry; the expressions of collagen type II and Aggrecan were determined with RT-PCR; and HE staining, toluidine blue staining, Safranin O staining of GAG, and collagen type II immunohistochemical staining were used for histological and immunohistochemical observations. Results At 28 days after incubation, all specimens formed disc tissue that was translucent and white with smooth surface and good elasticity in groups A and B; the specimens shrank into yellow spherical tissue without elasticity in group C. The wet weight and GAG content of specimens in groups A and B were significantly higher than those in group C (P lt; 0.05), but no significant difference was found between groups A and B in the wet weight (t=1.820 3, P=0.068 7) and in GAG content (t=1.861 4, P=0.062 7). In groups A and B, obvious expressions of collagen type II and Aggrecan mRNA could be detected by RT-PCR, but no obvious expressions were observed in group C; the expressions in groups A and B were significantly higher than those in group C (P lt; 0.05), but no significant difference was found between groups A and B in collagen type II mRNA expression (t=1.457 6, P=0.144 9) and Aggrecan mRNA expression (t=1.519 5, P=0.128 6). Mature cartilage lacunas and different degrees of dyeing for the extracellular matrix could be observed in groups A and B; no mature cartilage lacunas or collagen type II could be observed in group C. The expression of collagen type II around cartilage lacuna was observed in groups A and B, but no expression in group C; the gray values of groups A and B were significantly lower than that of group C (P lt; 0.01), but no significant difference was found between groups A and B (t=1.661 5, P=0.09 7 0). Conclusion Misshapen auricular chondrocytes from microtia can induce chondrogenic differentiation of human ADSCs in vitro.
To study how to repair the cartilage defect according to the principles of tissue engineering with acellular cartilage matrix as scaffold material. Methods The ear cartilage was obtained from a New Zealand white rabbit(weighing 2.4 kg )and then treated by a modified Courtman’s four-step method to produce the acellular cartilage matrix. Eighteen New Zealand white rabbits (aged 6 months, weighing 2.4-2.6 kg) with no sex l imit were divided into three groups. Forevery rabbit, two pieces of ear cartilage measured 1 cm × 1 cm were excised in each ear. Defects were repaired as follows: group A with the combined graft of acellular cartilage matrix and perichondium, group B with acellular cartilage matrix and group C with perichondium. Three animals in each group were killed 4 and 12 weeks postoperatively, respectively. Tissue samples obtained were analyzed with gross observation, hematoxyl in-eosin stain, Safranine O-alcian blue stain and type II collagen messenger RNA in situ hybridization respectively. Results In gross observation, the repaired sites in groups A and B were not change meaningfully in their shape 4 weeks postoperatively; but they felt a bit of thicker and harder 12 weeks postoperatively. In group C two repaired sites formed scabs at 2 weeks and perforated at 5 weeks. In histological observation, there was a sl ight inflammatory reaction surrounding the acellular cartilage matrix 4 weeks after it was implanted in groups A and B. The inflammatory cells were mainly lymphocytes. The perichondrium graft in group C was collapsed in the defects in HE stain. The defect sites were negative for Safranine O-alcian blue stain and type II collagen mRNA in situ hybridization in all groups. At 12 weeks cells were found in the acellular matrix which arranged in irregular manner in group A in HE stain and was positive for Safranine O-alcian blue stain and type II collagen mRNA in site hybridization. In groups B and C, no new cell was found in HE stain and the repaired sites were negative for Safranine O-alcian blue stain and type II collagen mRNA in situ hybridization. Conclusion Acellular
Objective To investigate the effectiveness of nasolabial flap and ear cartilage in repairing defects after nasal ala basal cell carcinoma resection. Methods Between January 2012 and August 2014, 8 patients with nasal ala basal cell carcinoma underwent tumor resection and defect repair with nasolabial flap and ear cartilage. Among the 8 patients, 5 were male and 3 were female, with an average age of 65 years (range, 45-76 years). The left side and right side were involved in 3 cases and 5 cases respectively. Carcinoma confirmed by pathological examination in all patients. The time between first biopsy and resection was 7-14 days (mean, 10 days). The defect ranged from 1.5 cm×1.5 cm to 2.0 cm×1.5 cm after tumor resection, and the size of nasolabial flaps ranged from 4.0 cm×1.5 cm to 5.0 cm×2.0 cm. The operations of cutting off the pedicle and thinning skin flap were performed at 6 months after first operation. Results All flaps survived. Incisions healed by first intention, and no related complication occurred. No carcinoma recurred after cutting off the pedicle. All patients were followed up for 6 months. All patients were satisfied with the nasal contour, symmetrical projection of the alar dome, and no obvious scar. Conclusion Nasolabial flap transfer and ear cartilage transplant method not only can repair the nasal ala defects, but also can avoid obvious scar and obtain good nasal ala contour profile. The shortcoming is that patients have to receive two operations.
ObjectiveTo explore the anthropometric changes of the auricle after auricular cartilage unfolding in moderate concha-type microtia patients, so as to provide the basis to help evaluate surgical timing and prognostic.MethodsA total of 33 children with moderate concha-type microtia, who were treated with auricular cartilage unfolding between October 2016 and September 2018 and met the inclusive criteria, were included in the study. There were 24 boys and 9 girls with an average age of 1.4 years (range, 1-3 years). Sixteen cases were left ears and 17 cases were right ears. The follow-up time was 12-23 months (mean, 17.5 months). The affected auricular detailed structures were observed and quantitatively analyzed before operation and at immediate after operation. The width, length, and perimeter of auricle before operation and at immediate after operation and at last follow-up were noted with three dimensional-scanning technology. The normal auricle was noted as control.ResultsThere were (7.5±1.0) and (11.3±0.8) structures of the affected auricle at pre- and post-operation, respectively, showing significant difference between pre- and post-operation (t=23.279, P=0.000). The length, width, and perimeter of the affected auricle constantly increased after operation, and there were significant differences between pre-operation and immediately after operation and between immediately after operation and last follow-up (P<0.05). The differences of length, width, and perimeter of the affected auricle between immediately after operation and last follow-up were (3.13±1.44), (2.44±0.92), and (8.50±3.76) mm, respectively. And the differences of length, width, and perimeter of the normal auricle between pre-operation and last follow-up were (3.16±1.54), (2.35±0.86), and (9.79±4.60) mm, respectively. There was no significant difference in the differences of length, width, and perimeter between the affected auricle and the normal auricle (P>0.05).ConclusionThe auricular cartilage unfolding in treatment of the moderate concha-type microtia can receive more ear structures and increase auricle sizes, which make it possible for free composite tissue transplantation. In addition, the affected and the contralateral normal auricles have a very similar growth rate and it offers the theoretical foundation for the early treatment for moderate concha-type microtia.