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Sustained plasmid DNA release from dissolving mineral coatings.

ABSTRACT: Calcium phosphate (CaP) minerals such as hydroxyapatite are able to bind a diverse range of biological molecules due to the presence of anions and cations in their crystal structure. The well-characterized ability of CaP minerals to bind and release plasmid DNA, coupled with the ability of biodegradable CaP coatings to form on the surface of common biomaterials, provides a potential mechanism for controlled release of plasmid DNA from various biomaterials. In this study we hypothesized that the release of plasmid DNA from CaP coatings formed on poly(lactide-co-glycolide) (PLG) substrates would be dependent on both the intrinsic properties of the CaP mineral coating and the surrounding solution conditions. Experiments were designed to consider two general parameters: (i) the stability of various CaP mineral coatings in solution environments that are relevant to physiological conditions; (ii) the relationship between mineral stability and sustained plasmid DNA release. Our results corroborate previous studies that have demonstrated a direct relationship between intrinsic mineral composition and mineral stability. In addition, we further demonstrate that ion composition and pH of the surrounding solution environment can significantly influence mineral stability. In turn, mineral stability significantly influenced release of plasmid DNA from mineral coatings in vitro, and the DNA release efficiency could be tuned by controlling the mineral properties in various solution environments. These CaP mineral coatings may be a useful platform for plasmid DNA delivery applications using various biomaterial platforms.

SUBMITTER: Choi S

PROVIDER: S-EPMC2902684 | biostudies-literature | 2010 Sep

REPOSITORIES: biostudies-literature

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Sustained plasmid DNA release from dissolving mineral coatings.

Choi Siyoung S Choi Siyoung S Murphy William L WL

Acta biomaterialia 20100318 9

Calcium phosphate (CaP) minerals such as hydroxyapatite are able to bind a diverse range of biological molecules due to the presence of anions and cations in their crystal structure. The well-characterized ability of CaP minerals to bind and release plasmid DNA, coupled with the ability of biodegradable CaP coatings to form on the surface of common biomaterials, provides a potential mechanism for controlled release of plasmid DNA from various biomaterials. In this study we hypothesized that the ...[more]

PMID: 20304109

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Project description:BackgroundMitochondrial DNA (mtDNA), a potent proinflammatory damage-associated molecular pattern, is released in large titers following trauma. The effect of trauma surgery on mtDNA concentration is unknown. We hypothesized that mtDNA and nuclear DNA (nDNA) levels would increase proportionately with the magnitude of surgery and both would then decrease rapidly.MethodsIn this prospective pilot, plasma was sampled from 35 trauma patients requiring orthopedic surgical intervention at six perioperative time points. Healthy control subjects (n = 20) were sampled. DNA was extracted, and the mtDNA and nDNA were assessed using quantitative polymerase chain reaction. Markers of cell necrosis were also assayed (creatine kinase, lactate dehydrogenase, and aspartate aminotransferase).ResultsThe free plasma mtDNA and nDNA levels (ng/mL) were increased in trauma patients compared with healthy controls at all time points (mtDNA: preoperative period, 108 [46-284]; postoperative period, 96 [29-200]; 7 hours postoperatively, 88 [43-178]; 24 hours, 79 [36-172]; 3 days, 136 [65-263]; 5 days, 166 [101-434] [healthy controls, 11 (5-19)]) (nDNA: preoperative period, 52 [25-130]; postoperative period, 100 [35-208]; 7 hours postoperatively, 75 [36-139]; 24 hours postoperatively, 85 [47-133]; 3 days, 79 [48-117]; 5 days, 99 [41-154] [healthy controls, 29 (16-54)]). Elevated DNA levels did not correlate with markers of cellular necrosis. mtDNA was significantly elevated compared with nDNA at preoperative period (p = 0.003), 3 days (p = 0.003), and 5 days (p = 0.0014). Preoperative mtDNA levels were greater with shorter time from injury to surgery (p = 0.0085). Postoperative mtDNA level negatively correlated with intraoperative crystalloid infusion (p = 0.0017). Major pelvic surgery (vs. minor) was associated with greater mtDNA release 5 days postoperatively (p < 0.05).ConclusionThis pilot of heterogeneous orthopedic trauma patients showed that the release of mtDNA and nDNA is sustained for 5 days following orthopedic trauma surgery. Postoperative, circulating DNA is not associated with markers of tissue necrosis but is associated with surgical invasiveness and is inversely related to intraoperative fluid administration. Sustained elevation of mtDNA levels could be of inflammatory origin and may contribute to postinjury dysfunctional inflammation.Level of evidenceProspective study, level III.

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Sustained plasmid DNA release from dissolving mineral coatings.

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Sustained plasmid DNA release from dissolving mineral coatings.

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