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Agricultural waste-based cellulose fibers have gained significant interest for a myriad of applications. Grewia optiva (G. optiva), a plant species, has been widely used for feeding animals, and the small branches’ bark is used for making rope. Herein, we have extracted cellulose fibers from the bark of G. optiva species via chemical treatments (including an alkaline treatment and bleaching). The gravimetric analysis revealed that the bark of G. Optiva contains cellulose (63.13%), hemicellulose (13.52%), lignin (15.13%), and wax (2.8%). Cellulose microfibre (CMF) has been synthesized from raw fibre via chemical treatment methods. The obtained cellulose fibers were crosslinked and employed as the matrix to encapsulate the bioactive plant extracts derived from the root of Catharanthus roseus (C. roseus). The microscopic images, XRD, FTIR, and antibacterial/antioxidant activity confirmed the encapsulation of natural extracts in the cellulose microfiber. The microscopic images revealed that the encapsulation of the natural extracts slightly increased the fiber’s diameter. The XRD pattern showed that the extracted cellulose microfiber had an average crystalline size of 2.53 nm with a crystalline index of 30.4% compared to the crystalline size of 2.49 nm with a crystalline index of 27.99% for the plant extract incorporated membrane. The water uptake efficiency of the synthesized membrane increased up to 250%. The antimicrobial activity of the composite (the CMF-E membrane) was studied via the zone inhibition against gram-positive and gram-negative bacteria, and the result indicated high antibacterial activity. This work highlighted G. optiva-derived cellulose microfiber as an optimum substrate for antimicrobial scaffolds. In addition, this paper first reports the antimicrobial/antioxidant behavior of the composite membrane of the C. roseus extract blended in the G. optiva microfiber. This work revealed the potential applications of CMF-E membranes for wound healing scaffolds.