Primary hepatocytes (PHCs) are widely used in various fields, but the progressive deterioration of liver-specific features in vitro significantly limits their application. While the transcriptional regulation and whole cell proteome (WCP) of PHCs have been extensively studied, only a small number of studies have addressed the role of posttranslational modifications in this process. To elucidate the underlying mechanisms that induce dedifferentiation, we carried out parallel quantifications of the transcriptome, WCP, ubiquitinome, and phosphoproteome of rat PHCs after 0, 6, 12, 24, and 48 h of in vitro culture. Our data constitute a detailed proteomic analysis of dedifferentiated PHCs including 2196 proteins, 2056 ubiquitinated sites, and 4932 phosphorylated peptides. We revealed a low correlation between the transcriptome and WCP during dedifferentiation. A combined analysis of the ubiquitinome with the corresponding WCP indicated that the dedifferentiation of PHCs led to an increase in nondegradative K27 ubiquitination. Functional analysis of the altered phosphoproteins suggested a significant enrichment in ferroptosis. In all, 404 proteins with both ubiquitination and phosphorylation were identified to be involved in critical metabolic events. Furthermore, Ptbp1, Hnrpd, Hnrnpu, and Srrm2 were identified as hub genes. Taken together, our data provide new insights into proteome dynamics during PHC dedifferentiation and potential targets to inhibit the dedifferentiation process.
Due to their capability of fabricating geometrically complex structures, additive manufacturing (AM) techniques have provided unprecedented opportunities to produce biodegradable metallic implants— especially using Mg alloys, which exhibit appropriate mechanical properties and outstanding biocompatibility. However, many challenges hinder the fabrication of AM-processed biodegradable Mg-based implants, such as the difficulty of Mg powder preparation, powder splash, and crack formation during the AM process. In the present work, the challenges of AM-processed Mg components are analyzed and solutions to these challenges are proposed. A novel Mg-based alloy (JDBM) powder with a smooth surface and good roundness was first synthesized successfully, and the AM parameters for Mg-based alloys were optimized. Based on the optimized parameters, porous JDBM scaffolds with three different architectures (biomimetic, diamond, and gyroid) were then fabricated by selective laser melting (SLM), and their mechanical properties and degradation behavior were evaluated. Finally, the gyroid scaffolds with the best performance were selected for dicalcium phosphate dihydrate (DCPD) coating treatment, which greatly suppressed the degradation rate and increased the cytocompatibility, indicating a promising prospect for clinical application as bone tissue engineering scaffolds.
Complicated and large acetabular bone defects present the main challenges and difficulty in the revision of total hip arthroplasty (THA). This study aimed to explore the advantages of three-dimensional (3D) printing technology in the reconstruction of such acetabular bone defects. We retrospectively analyzed the prognosis of four severe bone defects around the acetabulum in three patients who were treated using 3D printing technology. Reconstruction of bone defect by conventional methods was difficult in these patients. In this endeavor, we used radiographic methods, related computer software such as Materialise's interactive medical image control system and Siemens NX software, and actual surgical experience to estimate defect volume, prosthesis stability, and installation accuracy, respectively. Moreover, a Harris hip score was obtained to evaluate limb function. It was found that bone defects could be adequately reconstructed using a 3D printing prosthesis, and its stability was reliable. The Harris hip score indicated a very good functional recovery in all three patients. In conclusion, 3D printing technology had a good therapeutic effect on both complex and large bone defects in the revision of THA. It was able to achieve good curative effects in patients with large bone defects.