Abstract Background Platelet-derived Growth factors play key roles in tissue repair and regeneration, yet conventional platelet-rich plasma (PRP) formulations release these mediators inconsistently in vivo due to variability in platelet yield and activation dynamics. To overcome this limitation, direct administration of concentrated platelet-derived growth factor preparations has gained interest, though current manufacturing approaches for human platelet lysate (hPL), growth factor concentrates (GFC), and conditioned serum remain constrained by batch variability, incomplete platelet degranulation, and reliance on anticoagulants. Here, we examine alternative platelet activation workflows to establish a standardized, efficient, and reproducible method for high-yield growth factor recovery suitable for translational and clinical applications. Methods Nine GFC production protocols were compared, employing different combinations of freeze–thaw (FT) cycling, glass bead (GB) agitation, calcium (Ca 2 ) activation, and a novel Enriched Growth Factor (Enriched-GF) method. The objective was to identify a protocol capable of maximizing growth factor yield within a three-hour workflow. Optimal Ca 2 concentrations and GB conditions were determined from prior optimization studies and integrated into the Enriched-GF processing scheme. Platelet concentrates (n = 10 per protocol) were processed under each condition, and growth factor levels were quantified using ELISA. Results Growth factor yields differed significantly across protocols. The greatest and most consistent increases in growth factor release were observed with the Enriched-GF method combining GB activation, FT cycling, and Ca 2 stimulation. This approach resulted in markedly elevated concentrations of key regenerative mediators, including enhanced EGF release, a 4.5-fold increase in PDGF, maximal TGF-β liberation, and a four-fold increase in FGF2 relative to conventional platelet lysate or conditioned serum preparations. These results were reproducible across independent donor pools, demonstrating robustness and batch-to-batch consistency. Conclusion We describe a rapid and reproducible method for producing highly concentrated platelet-derived growth factors using a combined GB–FT–Ca 2 activation strategy. The Enriched-GF protocol consistently outperformed existing platelet lysate, conditioned serum, and conventional GFC preparation methods, yielding a standardized product with enhanced growth factor content. This Enriched-GF approach offers a clinically practicable solution for applications in regenerative medicine requiring reliable and high-yield growth factor delivery. Abstract Figure Schematic overview of platelet concentrate preparation from whole blood and the generation of different platelet lysates and growth factor-enriched serum using freeze-thaw, calcium gluconate, and glass bead activation methods.
Background Non-union is a common clinical complication that causes serious limb debilities and eventually leads to permanent limb impairments if not treated on time. This proof-of-concept study was undertaken to evaluate the osteogenic potential of autologous adipose derived stromal cells (ADSCs) in long bone non-unionadipose-derived stromal cells (ADSCs) in long-bone nonunion. Methods: Seven patients with long-standing atrophic non-union of long bones were treated with percutaneous implantation of a minimum of 100 million ADSC and platelet-rich plasma containing a minimum of 10 billion platelets into the non-union site of long bones under fluoroscopic guidance. Besides radiological imaging for evidence of consolidation, patients were also followed up for clinical parameters, standard lower extremity functional scale (LEFS) and SF12 scores. The patients were followed for a minimum of one year, & Intervention was considered a failure if no evidence of healing was observed up to 6 months post-procedure Results: We observed union in 6 of 7 (86%) patients within 4 months after the procedure. The first evidence of healing was visible within 8 weeks after treatment in five (71%) patients. Conclusions: The study demonstrated the efficacy of ADSCs as a better alternative to bone marrow aspirate concentrate (BMAC) in treating long bone non-union. Level of Evidence Level 3 Trial Registration: This study has been registered in the US Clinical Trial Registry (U.S. National Library of Medicine) with Trial registration no. [NCT04340284][1]. Titled as Adipose Derived Stromal Vascular Fraction (SVF) Application in treatment of Long Bone nonunion https://clinicaltrials.gov/ct2/show/[NCT04340284][1]?term=NCT04340284&draw=2&rank=1 Date of registration 09/04/2020 Keywords: Non-union, long bone fracture, Bone Marrow Aspirate Concentrate (BMAC), Adipose derived stromal cells (ADSCs), mesenchymal stem cells, Growth stimulators, Platelet Rich Plasma (PRP). ### Competing Interest Statement The authors have declared no competing interest. ### Clinical Trial NCT04340284 ### Funding Statement Funding is acknowledged from Anupam Hospital, Uttarakhand, India. (Grant no. 2012/02/ UK/ADM /023) ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: This trial was ethically approved by the Institutional Committee for Stem Cell Research and Therapy, Anupam Hospital, Uttarakhand, India. The trial is compliant with the consolidated standards of reporting trials (CONSORT). Informed prior consent was obtained from all the patients. This trial was registered in Clinicaltrials.gov-[NCT04340284][1]; Date of registration 09/04/2020; ClinicalTrial.gov under URL: https://clinicaltrials.gov/ct2/show/[NCT04340284][1]). I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable. Yes All data produced in the present study are available upon reasonable request to the authors [1]: /lookup/external-ref?link_type=CLINTRIALGOV&access_num=NCT04340284&atom=%2Fmedrxiv%2Fearly%2F2025%2F08%2F21%2F2025.07.17.25331624.atom
Introduction Acute patellar tendon injuries in children are rare but increasing due to more high-energy sports participation. These injuries often involve bony fractures, with isolated tendon avulsions being uncommon. Timely diagnosis and surgical intervention are essential to restore knee function and pre-injury activity levels. Case presentation A 12-year-old male student-athlete was brought to the hospital after his right knee buckled while playing football. He exhibited significant tenderness, swelling, and an inability to actively extend the knee. Clinically, a palpable gap below the inferior pole of the patella was detected. Discussion Initial X-ray imaging was inconclusive. MRI confirmed a proximal patellar tendon avulsion from the inferior patella, with a narrow soft tissue sleeve attached to the distally retracted tendon. No bony avulsion or was found. Interestingly, intraoperatively, there was a complete avulsion of the periosteal sleeve covering the patella. Transosseous suture repair with suture anchor augmentation was performed. Followed by full reattachment of the periosteal sleeve. Three years post-surgery, the patient became a professional basketball player with full knee motion, no pain or instability, and excelled in high-intensity activities without limitations. Conclusion Isolated proximal patellar tendon injury without bony avulsion is rare, and its association with complete periosteal sleeve detachment is exceptionally uncommon. The combination of transosseous repair and suture anchor augmentation ensured long-term stability. This approach effectively distributed tensile forces, minimizing the risk of re-injury and repair failure. A tailored rehabilitation ensured full knee recovery, with long-term follow-up confirming a return to professional sports without limitations.
