Lysahealالعربية

Science

The protocol, in detail

This page sets out the production protocol, the potency-release concept, and the protease-protection rationale behind Lysaheal, with a full reference list. It is written for clinicians, reviewers, and scientific partners who want more than the summary on the home page.

1. Source material and collection window

Lysaheal's feedstock is platelet units that have passed their 5–7 day transfusion shelf life and would otherwise be discarded. Units suspended in 100% plasma are prioritised because they yield higher fibrinogen content and roughly double gel yield compared with units in platelet additive solution. Our working hypothesis — not yet validated — is that a day 6–10 post-expiry collection window preserves adequate growth-factor content while maximising the volume of otherwise-wasted material available for use. An Icelandic blood-bank study found no difference between fresh and expired platelet concentrates for lysate production, which supports the general premise that expiry does not equal biological inactivity.

2. Pooling and lysis

Batches pool four units from at least 16 donors, which averages out donor-to-donor variability in growth-factor content — a known limitation of single-donor or autologous platelet-rich plasma. Lysis uses four freeze–thaw cycles (−80 °C / +37 °C) followed by centrifugation at 4,000 × g for 10 minutes, a protocol with precedent in the platelet-lysate literature used for cell-culture supplement production. Pathogen reduction is applied at the individual unit stage, using the pathogen-reduction system already in place at the host blood bank, before pooling.

3. Potency-based release

Rather than releasing batches on volume or platelet count alone, every batch is tested by ELISA for five growth factors — PDGF-BB, VEGF, TGF-β1, EGF and bFGF — and by a fibroblast scratch (migration) bioassay. A batch fails release if PDGF-BB or TGF-β1 falls below a pre-set specification. Dose is expressed in nanograms of growth factor per cm² of wound surface area, not millilitres of gel applied. To our knowledge, no existing commercial or blood-service platelet product states a measured growth-factor dose on this basis; this is a hypothesis about how the field should measure potency, not a validated standard.

4. Activation and application

10 mL of lysate is combined with 3.3 mL of 10% calcium gluconate and 0.4 mL of tranexamic acid, forming a fibrin gel within 30 minutes. This calcium-gluconate/tranexamic-acid activation avoids bovine thrombin and batroxobin, both used by some existing platelet-gel protocols, and has precedent in Italian blood-service practice (Vox Sanguinis, 2025). Reagent cost is under €3 per dose, and the protocol needs no capital equipment and no patient blood draw.

5. Protease protection and formulation

Chronic wounds are protease-rich environments: matrix metalloproteinase-9 (MMP-9) and neutrophil elastase degrade unprotected growth factors within hours of application, which is a plausible explanation for the inconsistent results seen with some topical growth-factor products. Lysaheal's fibrin matrix is designed to use heparin-binding retention to shield growth factors from this degradation. In parallel, we are developing a lyophilised (freeze-dried), single-use sachet formulation stabilised with trehalose, targeting stability at 45 °C to suit Saudi ambient conditions and, eventually, home application without a cold chain. Both the protease-protection mechanism and the 45 °C stability target are engineering hypotheses to be tested in the bench programme below, not demonstrated properties.

6. What is validated versus what is hypothesis

Validated in the published literature: the use of expired platelet units as a lysate source; the freeze–thaw lysis protocol; calcium-gluconate/tranexamic-acid activation; the retention of growth factors in lyophilised platelet-derived products over months of storage; and the protease pathology of chronic wounds. Still to be tested as Lysaheal-specific hypotheses: the day 6–10 post-expiry collection window; potency-based ng/cm² dosing as a superior release standard; the protease-protected formulation's real-world performance; 45 °C stability; and efficacy in sickle cell leg ulcers, a population with no prior platelet-therapy data of any kind.

7. Regulatory position

A pooled allogeneic topical product of this kind is likely to be classified by the Saudi Food and Drug Authority as a biological medicinal product. Lysaheal is engaging the SFDA early for classification guidance, and will point to the precedent of Italy's Ministerial Decree of 2 November 2015, which recognises platelet gel and platelet lysate as blood components for non-transfusional use when produced within transfusion services. No regulatory approval of any kind has been obtained for Lysaheal's product, and none is implied by this page.

References

  1. 1.Blood-component waste rate of 19.44% (2021–2023) and expiration as leading cause of discard (28.4%) at King Fahad Armed Forces Hospital, Saudi Arabia. [Citation — journal, year and DOI/URL to be confirmed.]
  2. 2.Average platelet disposal rate of 13% (range 6.7–25%) across 17 European countries. [Citation — journal, year and DOI/URL to be confirmed.]
  3. 3.Icelandic blood-bank study finding no difference between fresh and expired platelet concentrates for lysate production. [Citation — journal, year and DOI/URL to be confirmed.]
  4. 4.IDF Diabetes Atlas, 2024 — adult diabetes prevalence in Saudi Arabia of 23.1%. [DOI/URL to be confirmed.]
  5. 5.Systematic review of diabetic foot ulcer prevalence across five Arab countries, reporting 8.5% prevalence in Saudi Arabia (highest of the five). [Citation — journal, year and DOI/URL to be confirmed.]
  6. 6.Middle East meta-analysis reporting 33% amputation rate among diabetic foot ulcer patients, with Saudi rates significantly higher than regional peers. [Citation — journal, year and DOI/URL to be confirmed.]
  7. 7.Sickle cell disease prevalence of 134 per 1,000 population in Saudi Arabia's Eastern Province, and leg ulcer history in 8% of Saudi sickle cell patients (16.7% over age 50). [Citation — journal, year and DOI/URL to be confirmed.]
  8. 8.2020 treatment review identifying eighteen platelet-rich plasma studies in leg ulcers, none including sickle cell patients. [Citation — journal, year and DOI/URL to be confirmed.]
  9. 9.Li et al., "Allogeneic platelet-rich plasma for wound healing", Scientific Reports, 2024 — meta-analysis of 12 RCTs, 717 patients, RR 2.72 (95% CI 1.77–4.19). [DOI/URL to be confirmed.]
  10. 10.Acta Diabetologica, 2025 — meta-analysis of PRP vs. standard care in diabetic foot ulcers, 15 RCTs, 1,010 patients, RR 1.53. [Citation — full author list, title and DOI/URL to be confirmed.]
  11. 11.Piccin et al., cord blood platelet gel after revascularisation in chronic ulcers, 2018 — 79% vs. 46% ulcer area reduction vs. standard care. [DOI/URL to be confirmed.]
  12. 12.Vox Sanguinis, 2025 — Italian blood-service protocol for tranexamic-acid/calcium-gluconate platelet gel activation. [Citation — full author list and DOI/URL to be confirmed.]
  13. 13.Italian Ministerial Decree of 2 November 2015, recognising platelet gel and platelet lysate as blood components for non-transfusional use produced within transfusion services. [URL to be confirmed.]
  14. 14.International Working Group on the Diabetic Foot — historical position on platelet gel in diabetic foot ulcer guidelines. [Citation and URL to be confirmed.]
  15. 15.Retraction notice, diabetic foot PRP meta-analysis, 2025. [Citation and URL to be confirmed.]

Every statistic on this site is drawn from the sources above. Several citations are still missing full bibliographic detail (journal, volume, DOI) and are marked accordingly — see CONTENT_TODO.md in the project repository for the full list to complete before publication.