Histology and histopathology Vol.36, nº7 (2021)
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- PublicationOpen AccessSequential osseointegration from osseohealing to osseoremodeling - Histomorphological comparison of novel 3D porous and solid Ti-6Al-4V titanium implants(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2021) Frosch, Alice; Krohn, Sebastian; Buchhorn, Gottfried; Lehmann, Wolfgang; Frosch, Karl-Heinz; Füzesi, László; Frosch, StephanIn the present study, we analyzed the histological characteristics of osseointegration of an open-porous Ti-6Al-4V material that was produced in a space holder method creating a 3-D through-pores trabecular design that mimics the inhomogeneity and size relationships of trabecular bone in macro- as well as microstructure. Pairs of cylindrical implants with a porosity of 49% and an average pore diameter of 400 µm (PI) or equal sized solid, corundum blasted devices (SI) as reference were bilaterally implanted press fit in the lateral condyles of 16 rabbits. Histological examination was performed after 4 weeks of short-term osseohealing and 12 weeks of mid-term osseoremodeling and we summarized the criteria for sequential osseointegration. After 4 weeks, osteoid had already been largely replaced by mineralized woven bone in both types of implants but was only represented to a greater extent in the deeper pores of PI. The cortical as well as trabecular region showed regular osseohealing with excessive and spatially undirected formation of immature woven bone. A dense bone mass was found in the cortical area, while in the trabecular region the bone mass was reduced distinctly, presenting large lacuna-like recesses and a demarcating trabecular structure. The pores near the implant surface contained more mineralized woven bone than the deeper pores. After 12 weeks, the osseoremodeling was largely completed with a physiological maturation to lamellar bone. The newly formed bone mass increased for PI and SI compared to the 4-week group and osteoid was only detectable in the deeper pores. The inhomogeneous trabecular design of the pores enables an excellent ingrowth of mineralized lamellar bone after remodeling to a pore depth of 1800 µm, which proves a functional load transfer from the surrounding bone into the implant. According to the concept of osseointegration by Branemark and Albrektsson, the histological evaluation confirms a successful, superior osseointegration of the presented porous properties improving long-term implant stability. The presented study protocol allows an excellent evaluation and comparison of the sequential osseointegration from short-term osseohealing to midterm osseoremodeling.
- PublicationOpen AccessPersistent mdx diaphragm alterations are accompanied by increased expression and activity of calcium and muscle-specific proteins(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2021) Gaglianone, Rhayanna B.; Fonseca Bloise, Flavia; Lagrota-Candido, Jussara; Mermelstein, Claudia; Quirico-Santos, TherezaThe mdx mouse model of Duchenne Muscular Dystrophy (DMD) presents sarcolemma instability and develops a mild multi-stage dystrophinopathy characterized by intense myonecrosis with inflammatory infiltrate at 4-weeks; muscular regeneration at 12-weeks and persistent fibrosis onwards. Mdx diaphragm muscle has a more severe phenotype with structural and functional deterioration that closely resembles the diaphragm impairment responsible for DMD human patients' morbidity. Herein, we compared calcium deposits, activity of calciumrelated proteases, and expression of muscle-specific proteins in mdx diaphragm at 4-weeks and 12-weeks. We found increased calcium deposits mainly at 12- weeks, concomitant with high activity of calpains and matrix metalloprotease-9, but decreased expression of Myh4 (Myhc IIb) and Atp2a1 (SERCA1), and high expression of the myogenic regulatory factors Myod1 and Myog. Our results suggest that increased calcium deposits and persistent activity of calcium dependent proteases throughout the disease are involved in the degeneration and regeneration processes in the mdx diaphragm.
- PublicationOpen AccessA new practical classification of desmoplastic reaction in endoscopic forceps biopsy of colorectal cancer(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2021) Jing, Haiyan; Shang, Liang; Zhao, Shulei; Yao, Zhigang; Lv, Beibei; Zhu, XiaolongBackground. The histopathological discrepancy between endoscopic forceps biopsy (EFB) and post-resection specimens is considered a practical clinical problem. This retrospective study aimed to determine the current diagnostic concordance between the EFB and surgical specimens of colorectal cancer (CRC) and then investigated the useful factors in EFB diagnosis. Methods. We used the representative pathological data of 2188 CRCs. The comparison of histopathological discrepancy between EFB and the related surgical specimens was performed. Furthermore, 418 biopsy specimen slides in our hospital were reviewed to determine the classification of intratumor desmoplastic reaction (DR). Results. Among the 2188 patients, the positive sensitivity of EFB for adenocarcinoma was 82.7%. The discrepancy rate between the EFB and surgical specimens was 10.8-40.0% corresponding to different T stages. On the basis of DR classification, 32, 131, and 255 tumors were categorized as little, moderate and extensive, respectively. The correlation between DR classification and tumor invasion based on T stage was significant (Spearman's rho= 0.112; p<0.05). The extensive DR provided better estimates for advanced tumors than the little and moderate DR (χ2= 3.977, p=0.046). Besides DR, factors including deeper cutting the slides and histological types were significantly associated with "adenocarcinoma" diagnosis in EFB of CRCs (p<0.05). Conclusion. To the best of our knowledge, this is the first time that a DR classification specifically for EFB specimens was proposed. It might contribute to improve the accuracy of biopsy-based diagnosis of CRC.
- PublicationOpen Accessβ-Ecdysone attenuates cartilage damage in a mouse model of collagenase-induced osteoarthritis via mediating FOXO1/ADAMTS-4/5 signaling axis(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2021) Han, Junzhu; Guan, Jianzhong; Zhu, XunbingBackground. β-Ecdysone has been reported to perform a protective effect to prevent interleukin 1β (IL-1β)-induced apoptosis and inflammatory response in chondrocytes. In our study, the chondroprotective effects of β-Ecdysone were explored in a mouse model of collagenase-induced osteoarthritis (OA). Methods. Injection of collagenase in the left knee was implemented to establish a mouse model of OA. The histomorphological analysis was detected using safranine O staining. Serum pro-inflammatory cytokines were measured by ELISA assays. Protein expression in the femur and chondrocytes was analyzed using western blot. Chondrocyte apoptosis was evaluated by terminaldeoxynucleoitidyl transferase mediated nick end labeling (TUNEL) staining. Results. Treatment of OA mice with β-Ecdysone supplementation significantly inhibited the production of pro-inflammatory cytokines. Histologic examination exhibited that the degradation of proteoglycans and the loss of trabecular bone were observed in collagenaseinjected mice. However, OA-like changes were attenuated by β-Ecdysone administration in collagenaseinjected mice. Both in vivo and in vitro models, nuclear forkhead box O1 (FOXO1) protein expression was significantly reduced in the femur of collagenase-treated mice and IL-1β-stimulated chondrocytes. However, βEcdysone treatment was able to rescue FOXO1 protein expression in the nucleus to inhibit the transcription and translation of a disintegrin-like and metallopeptidase (reprolysin type) with thrombospondin type 1 motif, 4 (ADAMTS-4) and ADAMTS-5. Conclusion. The findings suggested that β-Ecdysone functioned as a FOXO1 activator to protect collagenaseinduced cartilage damage. FOXO1 might be a potential molecular target of β-Ecdysone for the effective prevention and treatment of OA.
- PublicationOpen AccessPatient iPSC-derived retinal organoids: Observable retinal diseases in-a-dish(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2021) Zhang, Xiao-Hui; Jin, Zi-BingInduced pluripotent stem cells (iPSCs), reprogrammed from human somatic cells, hold the capacity to differentiate into most human body cells. iPSCs can be differentiated into retinal organoids, a three-dimensional structured retina containing various retinal cells. Patient-specific retinal organoids provide a powerful disease model to recapitulate the disease to study the pathogenesis of inherited retinal dystrophies, to screen or discover new drugs, and most importantly to supply an unlimited cell source for retinal regeneration
- PublicationOpen AccessVisual deficits after traumatic brain injury(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2021) Rasiah, Pratheepa Kumari; Geier, Ben; Jha, Kumar Abhiram; Gangaraju, RajashekharTraumatic brain injury (TBI) is frequently described as any head injury ceasing the brain's normal function. Anatomically, developmentally, and physiologically, the eye is deemed as an extension of the brain. Vision in TBI is underrepresented, and the number of active clinical trials in this field are sparse. Frequently, visual problems are overlooked at the time of TBI, often resulting in progressive vision loss, lengthening, and impairing rehabilitation. TBI can be either penetrative or non-penetrative, associated with degeneration of neurons, apoptotic cell death, inflammation, microglial activation, hemorrhage associated with vascular dysfunction; however, precise animal modeling that mimics the extensive visual deficits of TBI pathology remain elusive. Recent works in both the diagnostics and therapeutics fields are starting to make substantial progress in the right direction. Discussion of current advancements in TBI animal models and the recent pathophysiological findings related to the neuro-glia-vascular unit (NVU) will help elucidate novel targets for potential lines of therapeutics. Only over the past decade have newer pharmaceutical and stem cell-based treatments begun to come to light. The potency for these new lines of TBI specific curatives will be discussed along with the review of current blast-induced TBI models, providing potential directions for future research.
- PublicationOpen AccessStroma composition and proliferative activity are related to therapy response in neoadjuvant treated pancreatic ductal adenocarcinoma(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2021) Haeberle, Lena; Insilla, Andrea Cacciato; Kap, Anne-Christine; Steiger, Katja; Schlitter, Anna Melissa; Konukiewitz, Björn; Demir, Ihsan Ekin; Friess, Helmut; Esposito, IreneBackground. Tumor regression grading (TRG) based on histopathology is the main tool to assess therapy effects after neoadjuvant therapy (NAT) of pancreatic ductal adenocarcinoma (PDAC). However, reliable markers to distinguish therapy effects from preexisting tumor features are lacking. The aim of this study was the characterization of PDAC after NAT, focusing on the stroma. Material and Methods. Tissue samples from patients resected for PDAC after NAT (n=27) were analyzed. TRG was assessed using the Royal North Shore (RNS) system. Stromal composition was evaluated by Movat's stain. Immunohistochemistry (IH) for Ki-67 and five previously established stroma markers (alpha-Crystallin B, alpha-Smooth muscle actin (alpha-SMA), Neurotrophin-3 (NT-3), SPARC and Tenascin C) was also performed. Results were compared with therapynaïve PDACs (n=10). Results. Most cases showed a moderate response (RNS 2; 74%), while 15% displayed a poor response (RNS 3), and 11% a good response (RNS 1). No complete response was observed. Poor regression was associated with mucin-rich stroma, while good regression was associated with collagen-rich stroma. Cases with poorer therapy response had significantly higher proliferation. Higher peritumoral staining intensity for alpha-SMA and Tenascin C also showed a trend towards an association with poor regression. Conclusions. Similar to the stroma in therapy-naïve PDAC, the stroma of PDAC after NAT is heterogeneous. Distinguishing between desmoplastic stroma and therapy-induced fibrosis by single markers is not possible. Movat's pentachrome stain, IH for Ki-67, and to some extent for Tenascin C and alpha-SMA, can help detect poor histopathological response to NAT.
- PublicationOpen AccessC-C motif chemokine ligand 14 inhibited colon cancer cell proliferation and invasion through suppressing M2 polarization of tumor-associated macrophages(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2021) Li, Na; Liang, Xiao; Li, Jiawen; Li, Teng; Guo, ZuomingBackground. Colon cancer is one of the most common cancers with high incidence and high mortality. Chemokines play a crucial role in the development of cancer. Methods. Here, qRT-PCR was performed to detect gene expression. Western blot and immunohistochemistry were implemented to examine the expression of C-C motif chemokine ligand 14 (CCL14) in colon tumors. Besides, the expression of CD68 and CD206 in tumors was measured by immunohistochemistry. The percentages of M1- and M2-polarized macrophages were detected by flow cytometry. Furthermore, CCK-8 assay was performed to detect cell proliferation, and Transwell assay for cell invasion. Results. CCL14 was decreased in both colon tumors and colon cancer cells, and many tumor-associated macrophages (TAMs) infiltrated into the tumor. An increase CCL14 inhibited colon cancer cell proliferation. Importantly, CCL14 promoted THP-1 to M1 polarization induced by LPS and IFN-γ, and inhibited THP-1 to M2 polarization induced by IL-4 and IL-13. Besides, CCL14 enhanced the inhibition of M1-polarized macrophages to colon cancer cell proliferation and invasion, and reversed the promotion of M2-polarized macrophages to cell proliferation and invasion. Conclusion. Our data demonstrated that CCL14 inhibited the proliferation and invasion of colon cancer cells through suppressing the formation of M2-like TAMs
- PublicationOpen AccessBiological and biocompatible characteristics of fullerenols nanomaterials for tissue engineering(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2021) Zhao, Yizhe; Shen, Xinyuan; Ma, Ruimeng; Hou, Yiting; Qian, Yun; Fan, Cunyi. Fullerenes, as hydrophobic molecules, are limited in biomedical function due to their very low solubility. But taking C60(OH)x as an example, the properties of fullerenols were analyzed. It was found that fullerenols had good stability, water solubility, good biocompatibility and low cytotoxicity by adding a hydroxyl group to carbon atoms. In the biomedical field, it has been found that fullerene C60 can be used as a powerful free radical scavenger, with antioxidant activity, with antibacterial and inhibitory effects on cancer cells. Fullerenols inherit the good properties of fullerenes, and are better used in cancer treatment, including loading drug therapy and directly as an anticancer drug. In addition, fullerenols are also used in the repair of myocardial injury, the treatment of myocardial infarction and neuroprotection. With the development of tissue engineering technology, the preparation of nerve scaffolds which can improve ischemia, hypoxia and oxidative stress after nerve injury has become a research hotspot. The electron absorption and reduction characteristics of fullerenols in biomedical research bring new ideas for the treatment of oxidative stress in the repair of peripheral nerve defects. It seems that the research on fullerenols loaded neural scaffold has great prospects.