ObjectiveTo summarize the treatment strategies and clinical experiences of 5 cases of giant plexiform neurofibromas (PNF) involving the head, face, and neck. MethodsBetween April 2021 and May 2023, 5 patients with giant PNFs involving the head, face, and neck were treated, including 1 male and 4 females, aged 6-54 years (mean, 22.4 years). All tumors showed progressive enlargement, involving multiple regions such as the maxillofacial area, ear, and neck, significantly impacting facial appearance. Among them, 3 cases involved tumor infiltration into deep tissues, affecting development, while 4 cases were accompanied by hearing loss. Imaging studies revealed that all 5 tumors predominantly exhibited an invasive growth pattern, in which 2 and 1 also presenting superficial and displacing pattern, respectively. The surgical procedure followed a step-by-step precision treatment strategy based on aesthetic units, rather than simply aiming for maximal tumor resection in a single operation. Routine preoperative embolization of the tumor-feeding vessels was performed to reduce bleeding risk, followed by tumor resection combined with reconstructive surgery. Results All 5 patients underwent 1-3 preoperative embolization procedures, with no intraoperative hemorrhagic complications reported. Four patients required intraoperative blood transfusion. A total of 10 surgical procedures were performed across the 5 patients. One patient experienced early postoperative flap margin necrosis due to ligation for hemostasis; however, the incisions in the remaining patients healed without complications. All patients were followed up for a period ranging from 6 to 36 months, with a mean follow-up duration of 21.6 months. No significant tumor recurrence was observed during the follow-up period. Conclusion For patients with giant PNF involving the head, face, and neck, precision treatment strategy can effectively control surgical risks and improve the standard of aesthetic reconstruction. This approach enhances overall treatment outcomes by minimizing complications and optimizing functional and cosmetic results.
【摘要】 目的 探討微弧氧化（microarc oxidation，MAO）結合應用于純鈦種植體表面處理的可行性。 方法 根據對純鈦鈦片處理的不同將實驗分為對照組（A組，不作處理）、MAO組（B組，純鈦片上進行MAO處理）及MAO加Ⅰ型膠原組（C組，純鈦片上MAO處理后吸附Ⅰ型膠原）。將成骨細胞培養于各組鈦片上，通過掃描電鏡、MTT法檢測不同時間點各組鈦片表面的細胞生長及增殖情況，并檢測堿性磷酸酶（alkaline phosphatase，ALP）活性。 結果 掃描電鏡顯示成骨細胞在C組鈦片上細胞黏附情況優于A、B組；MTT法及ALP活性檢測示培養3、6 d，成骨細胞在C組鈦片上的增殖及ALP活性與A、B組比較差異均有統計學意義（Plt;0.05）。 結論 MAO結合Ⅰ型膠原處理的鈦片可更有效提高成骨細胞表面附著、增殖，且具有較高的ALP活性。【Abstract】 Objective To study the feasibility of applying microarc oxidation (MAO) with collagen Ⅰ in surface modification of pure titanium. Methods According to different processing methods, the pure titanium was divided into three groups: the control group (without surface modification, group A), MAO group (with microarc oxidation applied in pure titanium surface modification, group B), MAO+Ⅰ group (with microarc oxidation and collagen Ⅰ applied in pure titanium treatment, group C). Osteoblasts were cultured on the surface of titanium in each group, and the cell proliferation in each group was detected at different time points by scanning electron microscopy and MTT method. Moreover, the activity of alkaline phosphatase (ALP) was also detected. Results Scanning electron microscopy showed that adhesion of osteoblasts for group C was better than group A and group B. MTT method and ALP activity detection indicated that there was a significant difference between group C and group A, B in cell proliferation and ALP activity on the third and sixth day of cultivation (Plt;0.05). Conclusion MAO with collagen Ⅰ applied in surface modification of pure titanium may increase osteoblast attachment, and promote its proliferation and ALP activity.
To investigate the inhibitory effect of Col I A1 antisense ol igodeoxyneucleotide (ASODN) transfection mediated by cationic l iposome on Col I A1 expression in human hypertrophic scar fibroblasts. Methods Scar tissue was obtained from volunteer donor. Human hypertrophic scar fibroblasts were cultured by tissue block method. The cells at passage 4 were seeded in a 6 well cell culture plate at 32.25 × 104 cells/well, and then divided into 4 groups: group A, l iposomeand Col I A1 ASODN; group B, Col I A1 ASODN; group C, l iposome; group D, blank control. At 8 hours, 1, 2, 3 and 4 days after transfection, total RNA of the cells were extracted, the expression level of Col I A1 mRNA was detected by RT-PCR, the Col I A1 protein in ECM was extracted by pepsin-digestion method, its concentration was detected by ELISA method. Results Agarose gel electrophoresis detection of ampl ified products showed clear bands without occurrence of indistinct band, obvious primer dimmer and tailing phenomenon. Relative expression level of Col I A1 mRNA: at 8 hours after transfection, group A was less than groups B, C and D (P lt; 0.05), and groups B and C were less than group D (P lt; 0.05), and no significant difference was evident between group B and group C (Pgt; 0.05); at 1 day after transfection, groups A and B were less than groups C and D (P lt; 0.05), and there was no significant difference between group A and group B, and between group C and group D (P gt; 0.05 ); at 2 days after transfection, there were significant differences among four groups (P lt; 0.05); at 3 and 4 days after transfection, group A was less than groups B, C and D (P lt; 0.05), group B was less than groups C and D (P lt; 0.05), and no significant difference was evident between group C and group D (P gt; 0.05). Concentration of Col I protein: at 8 hours after transfection, group A was less than groups B, C and D (P lt; 0.05), groups B and C were less than group D (P lt; 0.05), and no significant difference was evident between group B and group C (P gt; 0.05); at 1 day after transfection, significant differences were evident among four groups (P lt; 0.05); at 2, 3 and 4 days after tranfection, groups A and B were less than groups C and D (P lt; 0.05), and no significant difference was evident between group A and group B (P gt; 0.05). Conclusion Col I A1 ASODN can inhibit mRNA and protein expression level of Col I A1. Cationic l iposome, as the carrier, can enhance the inhibition by facil itating the entry of ASODN into cells and introducing ASODN into cell nucleus.
ObjectiveTo explore the interaction between immune cell infiltration and extracellular matrix (ECM) in diffuse gastric cancer (DGC), and to identify novel diagnostic biomarkers and therapeutic targets. MethodsTranscriptomic data of DGC patients from The Cancer Genome Atlas (TCGA) database were analyzed to screen potential regulator factor of immune-related and ECM receptor-related signaling pathways. Differential expression of the identified regulator was assessed between the DGC tissues and the adjacent gastric tissues. Bioinformatics analysis was utilized to evaluate the relation between the regulator factor and immune cell infiltration and ECM, as well as prognosis. The clinical validation was performed using 90 paraffin-embedded DGC tissues and adjacent gastric tissues from the patients treated at The Lanzhou University Second Hospital (hereafter “our hospital”) from January 2017 to December 2019. The immunohistochemical staining was employed to examine the expression of regulator factor, followed by analysis of its association with immune cell infiltration, clinicopathologic features, and prognosis. Additionally, 10 paired DGC tissues and adjacent gastric tissues from the patients treated in our hospital in 2024 were collected for validation using real-time quantitative PCR to assess mRNA expression. The significance level was set at α=0.05. ResultsThe collagen type I alpha 1 chain (COL1A1), a potential regulator factor linked to immune and ECM receptor signaling pathways, was identified from the TCGA database. The COL1A1 was significantly overexpressed in the DGC tissues compared to the adjacent gastric tissues (P<0.001), and its high expression correlated with poorer prognosis [HR(95%CI)=2.98(1.21, 7.30), P=0.017]. The COL1A1 gene expression negatively correlated with CD8+ T cell enrichment score (CIBERSORT: r=?0.17, P<0.001; xCELL: r=?0.32, P<0.001) but positively correlated with M2 tumor-associated macrophage enrichment score (CIBERSORT: r=0.32, P<0.001; xCELL: r=0.24, P<0.001). The clinical validation confirmed that the COL1A1 protein and mRNA were both overexpressed in the DGC tissues (P<0.001). The patients with high COL1A1 protein expression had worse overall survival (P<0.001), and high expression (vs. low) was an independent risk factor for postoperative overall survival [HR(95%CI)=6.607(3.374, 12.940), P<0.001]. The COL1A1 protein expression positively correlated with CD163 (an M2 macrophage marker; r=0.76, P<0.001) and negatively with CD8+ (T cell marker, r=?0.84, P<0.001). ConclusionThis study demonstrates that COL1A1 is a potential therapeutic target for immune suppression and ECM interaction in DGC and a critical prognostic factor for long-term survival in patients with DGC.
Objective
To investigate the effect of collagen type I concentration on the physical and chemical properties of the collagen hydrogel, and to analyze the effect of different concentrations of collagen type I hydrogel on the phenotype and gene expression of the chondrocytes in vitro.
Methods
Three kinds of collagen hydrogels with concentrations of 12, 8, and 6 mg/ mL (C12, C8, and C6) were prepared, respectively. The micro-structure, compressive modulus, and swelling ratio of the hydrogels were measured and analyzed. The chondrocytes at 2nd passage were cocultured with three kinds of collagen hydrogels in vitro, respectively. After 1-day culture, the samples were stained with fluorescein diacetate (FDA) / propidium iodide (PI) and the cell activity was observed under confocal laser microscope. After 14-day culture, HE staining and toluidine blue staining were carried out to observe the histological morphology, and mRNA expressions of chondrocytes related genes (collagen type II, Aggrecan, collagen type I, collagen type X, Sox9) were determined by real-time fluorescent quantitative PCR.
Results
With the increase of collagen type I concentration from 6 to 12 mg/mL, the physical and chemical properties of the collagen hydrogels changed significantly: the fiber network became dense; the swelling ratios of C6, C8, and C12 were 0.260 ± 0.055, 0.358 ± 0.072, and 0.539 ± 0.033 at 192 hours, respectively, showing significant differences among 3 groups (P lt; 0.05); and the compression modulus were (4.86 ± 0.96), (7.09 ± 2.33), and (11.08 ± 3.18) kPa, respectively, showing significant differences among 3 groups (P lt; 0.05). After stained with FDA/PI, most cells were stained green, and few were stained red. The histological observation results showed that the chondrocytes in C12 hydrogels aggregated obviously with b heterochromia, chondrocytes in C8 hydrogels aggregated partly with obvious heterochromia, and chondrcytes in C6 hydrogels uniformly distributed with weak heterochromia. Real-time fluorescent quantitative PCR results showed that the mRNA expressions of collagen type II and Aggrecan were at the same level in C12, C8, and C6; the expressions of collagen type I, Sox9, and collagen type X were up-regulated with the increase of collagen type I hydrogels concentration, and the expressions were the highest at 12 mg/mL and were the lowest at 6 mg/mL, showing significant differences among 3 groups (P lt; 0.05).
Conclusion
Increasing the concentration of collagen hydrogels leads to better mechanical properties and higher shrink-resistance, but it may induce the up-regulation of cartilage fibrosis and hypertrophy related gene expression.
ObjectiveTo evaluate the effectiveness of Cross-Union surgery for the treatment of pseudarthrosis of the tibia (PT) with neurofibromatosis type 1 (NF1). MethodsThe clinical data of 8 children of PT with NF1 who met the selection criteria between January 2018 and December 2023 was retrospectively analyzed. There were 5 boys and 3 girls, and the operative age ranged from 1.8 to 13.3 years with a median age of 3.5 years. According to Paley classification, there were 2 cases of type 2a, 2 cases of type 3, 2 cases of type 4a, and 2 cases of type 4c. There were 5 cases of first operation and 3 cases of re-fracture after previous operation. Six cases had leg length discrepancy before operation, and 2 of them had shortening over 2.0 cm. Except for 1 case of ankle fusion, the other 7 cases had ankle valgus. Preoperative coronal/sagittal angulation was recorded. Postoperative pseudarthrosis healing and refracture were observed. Leg length discrepancy and tibiotalar angle were measured and recorded before operation and at last follow-up. Inan imaging evaluation criteria was used to evaluate the imaging effect. ResultsAll patients were followed up 12-37 months (mean, 23.5 months). One pseudarthrosis failed to heal at 12 months after operation and healed at 3 months after reoperation, while the other pseudarthrosis healed with a healing rate of 87.5% and a healing time of 4-8 months (mean, 5.3 months). No refracture occurred during the follow-up. At last follow-up, there were 2 new cases with leg length discrepancy, which were 0.7 cm and 1.3 cm, respectively. In 2 cases with the leg length discrepancy more than 2.0 cm before operation, the improvement was from 4.1 cm and 12.6 cm to 2.1 cm and 9.0 cm, respectively. There was no significant difference in leg length discrepancy between pre- and post-operation in 8 cases (P>0.05). At last follow-up, 6 patients still had ankle valgus, and there was no significant difference in the tibiotalar angle between pre- and post-operation (P>0.05); the tibial coronal/sagittal angulation significantly improved when compared with that before operation (P<0.05). According to Inan imaging evaluation criteria, 1 case was good, 6 cases were fair, and 1 case was poor. Conclusion Cross-Union surgery is an effective method for the treatment of PT with NF1 in children, can achieve good bone healing results with a low risk of re-fracture. The surgery may not have significant effects on leg length discrepancy and ankle valgus, and further treatment may be required.
Objective To investigate the effect of vanadate on prol iferation and collagen type I production of rat medial collateral l igament (MCL)fibroblasts. Methods A total of 12 adult male SD rats were included, weighing 350-375 g. MCL was cut into small pieces (1 mm × 1 mm × 1 mm) in aseptic conditions, and then placed and cultured in culture chambers. Fibroblasts were passaged with 0.25% trypsin. The vanadate (0, 1.0, 2.5, 5.0 ng/mL) was added in the 3rd passage MCL fibroblasts, respectively, and the samples were divided into 4 groups (0, 1.0, 2.5, 5.0 ng/mL groups). MTT was used to measure the cell prol iferation. The production of collagen type I was measured by RT-PCR and ELISA. Twelve samples in each group were measured. Results In fibroblast prol iferation, the absorbency values of the 1.0, 2.5, 5.0 ng/mL groups were significantly different from that of the 0 ng/mL group (P lt; 0.05). The absorbency values of the 0, 1.0, 2.5, and 5.0 ng/mL groups were 0.213 ± 0.016, 0.327 ± 0.023, 0.449 ± 0.137, and 0.561 ± 0.028, respectively. In collagen secretion, vanadate in 1.0, 2.5, 5.0 ng/mL groups could significantly induce the production of collagen type I (P lt; 0.05) ina dose-dependent manner. The expressions of collagen type I of 0-5 ng/mL groups were 0.47 ± 0.02, 0.51 ± 0.03, 0.60 ± 0.01, and 0.72 ± 0.02, respectively. There was significant difference between the 1.0, 2.5, 5.0 ng/mL groups and 0 ng mL group (P lt; 0.05). RT-PCR displayed a dramatic increase of band density. The ratio of band density in the 0-5 ng mLgroups was 1.37 ± 0.76, 1.97 ± 0.53, 2.41 ± 0.94, and 2.73 ± 0.82, respectively. The gene expression of collagen type I in the 1.0, 2.5 and 5.0 ng/mL groups was higher than that in the 0 ng/mL group, and there was significant difference (P lt;0.05). There were statistical significant differences among 1.0, 2.5 and 5.0 ng/mL groups in each index mentioned above.Conclusion Vanadate can effectively induce the prol iferation of fibroblasts and the production of collagen type I in vitro, which may provide a new approach to the treatment of MCL injury.
Objective To investigate the possibility of repairing articular cartilage defects with the mesenchymal stem cells(MSCs) seeded type Ⅰ collagen-glycosaminoglycan(CG) matrices after being cultured with the chondrogenic differentiation medium. Methods The adherent population of MSCs from bone marrow of10 adult dogs were expanded in number to the 3rd passage. MSCs were seeded intothe dehydrothermal treatment (DHT) crosslinked CG matrices; 2×106 cells per 9mm diameter samples were taken. Chondrogenic differentiation was achieved by the induction media for 3 weeks. Cell contractility was evaluated by the measuement of the cell-mediated contraction of the CG matrices with time inculture.The in vitro formation of the cartilage was assessed by an assayemploying immunohistochemical identification of type Ⅱ collagen and by immunohistochemistry to demonstrate smooth muscle actin （SMA）. The cells seededingCGs wereimplanted into cartilage defectsof canine knee joints. Twelve weeks after surgery, the dogs were sacrificed and results were observed. Results There was significant contraction of the MSCsseeded DHT crosslinked CG scaffolds cultured in the cartilage induction medium. After 21 days, the MSCseeded DHT crosslinked matrices were contracted to 64.4%±0.3%; histologically, the pores were found to be compressedandthe contraction coupled with the newly synthesized matrix, transforming the MSCsseeded CG matrix into a solid tissue in most areas. The type Ⅱ collagen staining was positive. The SMA staining was positive when these MSCs were seeded and the contracted CGs were implanted into the cartilage defects of the canine knee joints to repair the cartilage defects. The function of the knee joints recovered and the solid cartilaginous tissue filled the cartilage defects. Conclusion The results demonstrates that MSCs grown in the CG matrices can produce a solid cartilaginous tissuecontaining type Ⅱ collagen after being cultured with the chondrogenic differentiation medium and implanted into cartilage defects. We hypothesize that the following steps can be performed in the chondrogenic process: ①MSCs express SMA, resulting in matrix contraction, thus achieving a required cell density (allowing the cells to operate in a necessary society)； ②Cells interact to form a type Ⅱ collagencontaining extracellular matrix (and cartilaginous tissue)； ③Other factors, suchas an applied mechanical stress, may be required to form a mature cartilage with the normal architecture.
