Acne vulgaris, caused by the bacterium P. acnes, is the most common cutaneous disorder in adolescents. Acne is not a life-threatening condition, however, it has significant psychological and physical effects on patients. P. acnes is capable of causing remarkable inflammatory responses. Topical therapy is advisable in acne treatment in mild to moderate acne conditions. Recently, available treatments for acne are mostly based on antibiotics and retinoids. The use of antibiotics has the limitations of developing bacterial resistance, while retinoids are highly teratogenic. The bacterial resistance and teratogenicity of isotretinoin led scientists to search for more potent and safer therapeutic options. Camel milk has been used in the preparation of pharmaceutical and cosmetic compositions such as anti-acne products. It was found that camel milk composition is closer to the human milk. Passive immunity is provided to new-borns by Immunoglobulins present in colostrum until its own immune system matures. The concentration in colostrum of specific antibodies against pathogens can be raised by immunizing a mammal with these pathogens or their antigens. Immunized milk products are preparations made of such hyper-immune antibodies enriched from it. Camel milk and milk-derived products have a growing potential therapeutic value. The objective of this research is to evaluate the antimicrobial and anti-inflammatory effects of immunized camel milk whey and its components against P. acnes and to develop a formula that harnesses all its beneficial properties. P. acnes was heat-killed in culture to obtain their outer membrane proteins (OMPs). The OMPs were mapped and quantified using SDS-PAGE and Bradford Assay, respectively. OMPs were used to prepare a vaccination injected into female lactating camels in timely manner. The immunized camel milk was collected then pasteurized, delipidized, de-caseinated, and freeze-dried to obtain the whey into powder form. The anti-P.acnes antibodies in the immunized camel whey were separated using Protein G and Protein A Chromatography and quantified by SDS-PAGE and ELISA, respectively. Then, the antimicrobial peptides (AMPs) including PGRPs and Lactoferrin were isolated by Heparin Affinity Chromatography, and mapped by SDS-PAGE. The antimicrobial and anti-inflammatory activity of the immunized camel whey and its constituents against P. acnes was evaluated in vitro. In vivo activity was also evaluated using rabbit ear model. After the activity of the immunized camel whey was confirmed, four topical formulae were developed and their composition and preparation protocol were patented. The findings of this research imply that the constituents of immunized camel milk whey (i.e. polyclonal anti-P. acnes antibodies, peptidoglycan recognition proteins (PGRPs), and lactoferrin) possess significant antimicrobial and anti-inflammatory activity against P. acnes, 90% and 70%, respectively. Topical formulae including; facial wash, serum, cream and gel enriched with 3% to 5% of immunized camel milk whey were developed and their activity against P. acnes was evaluated. Immunized camel milk whey that has been developed in this research exerts significant antimicrobial and anti-inflammatory activity against P. acnes and the unique composition formulated has been patented. Furthermore, the stability of the topical cream was validated to show that the antibodies in the cream formula retained around 75% of their activity.
|Date of Award||11 Jun 2020|
- University Of Strathclyde
|Supervisor||Ibrahim Khadra (Supervisor) & Alexander Mullen (Supervisor)|