bioRxiv Science⌕ Search

Biology subjects

Pacheco, Y. C.

Publications and source records attributed to Pacheco, Y. C..

2 recordsLinked to original sources

A Feature Learning Model Identifies Predictive Attributes of Mesenchymal Stromal Cell Efficacy

The therapeutic efficacy of human mesenchymal stromal cells (hMSCs) is highly variable, limiting their clinical translation for musculoskeletal diseases and other regenerative medicine applications. There is a poor understanding of the critical quality attributes correlating to therapeutic efficacy of hMSCs. To address this challenge, we analyzed pre-clinical in vitro secretome profiles and in vivo therapeutic efficacy of hMSCs from multiple human donors. hMSCs from different donors showed significant differences between donors in therapeutic efficacy when assessed in a rat post-traumatic osteoarthritis (OA) model. A partial least squares feature learning model was trained to evaluate differences between more and less therapeutic donor hMSCs by examining cytokine secretion profiles, to predict donor-specific therapeutic outcomes. More therapeutic hMSCs exhibited increased secretion of GM-CSF, GRO, IL-4, and PDGF-AA, whereas less therapeutic donors had higher TNF-, IL-6, and MCP-1 secretion. The cytokine profile was accompanied by evaluation of MAPK pathway, which revealed distinct differences in phospho-protein signaling between more and less therapeutic hMSC secretome profiles. Pharmacological inhibition of JNK signaling in more therapeutic donor cells decreased hMSC secretion of the key therapeutic associated cytokines and shifted hMSC secretome towards a less therapeutic profile. Prospective validation of cells from additional donors demonstrated significant correlations between predicted and observed pre-clinical in vivo efficacy to attenuate OA. This approach identifies critical quality attributes enabling consistent prediction of therapeutic potency, thereby addressing a major barrier to scalable and effective cell therapies. These findings advance precision cell-based therapies and offer a framework for standardized donor screening in clinical applications. SummaryA feature learning model was developed, trained, and validated to identify critical quality attributes of MSCs that predict therapeutic potency.

bioengineering↗

Melt Electrowritten Scaffold-Reinforced Affibody-Conjugated Hydrogels for Controlled Bone Morphogenetic Protein-2 Delivery

Bone morphogenetic protein-2 (BMP-2) is clinically used to promote bone regeneration but suffers from uncontrolled release when delivered from collagen sponges, necessitating high doses that can cause adverse effects. Hydrogels offer tunable protein release but are limited by weak mechanics and poor stability during storage and handling. Here, we introduce a two-part protein delivery platform that integrates mechanical reinforcement with affinity-controlled protein release. We developed a melt electrowritten (MEW) scaffold-reinforced, affibody-conjugated polyethylene glycol maleimide (PEG-mal) hydrogel for affinity-controlled BMP-2 delivery. MEW scaffolds improved hydrogel handling, compressive resistance, and stability during lyophilization and rehydration, without altering bulk stiffness. Engineered BMP-2-specific affibodies provided affinity-based control over BMP-2 release. This ability to control BMP-2 release was preserved after lyophilization and rehydration of the hydrogels. In vivo, affibody conjugation of high-affinity affibodies to the hydrogels significantly enhanced BMP-2 retention in subcutaneous implants, while MEW reinforcement significantly increased bone volume and defect bridging in rat femoral bone defects. This affibody-conjugated, MEW scaffold-reinforced hydrogel system effectively integrates mechanical reinforcement with tunable protein-material affinity interactions, advancing hydrogel-based delivery strategies for BMP-2 and other protein therapeutics in musculoskeletal repair.

bioengineering↗