Nanomaterials have been used for various applications in recent years due to their interesting physiochemical properties. Nevertheless, conventional physical and chemical ways of synthesizing nanomaterials are expensive and easily result in hazardous byproducts that are hazardous to the environment and public health. One of the very prominent applications of bio-mediated nanomaterials is in the field of healthcare applications. These nanomaterials are ideal candidates for drug delivery, imaging, and therapeutic applications. Additionally, bio-mediated nanomaterials are promising in antimicrobial therapy. Bacterial cellulose (BC) shows every characteristic of a biopolymer for healthcare applications, having a high surface area-to-volume ratio, tunable surface chemistry, great water-holding capacity, biocompatibility and biodegradability. The biosynthesis of BC using Komagataeibacter xylinus is very cost-effective and eco-friendly. BC itself is a very good biopolymer with wound-healing properties. There are various probiotic bacteria that have beneficial effects on our health, such as several Lactobacillus sp. have antimicrobial properties against pathogenic bacteria; Pediococcus pentosaceus have antimicrobial, antioxidant and antitumor activity. Combining the physiochemical properties of BC as a biopolymer and the medicinal properties of probiotic bacteria, probiotic bacterial cellulose (PBC) can be derived as a bio-mediated nanomaterial for different healthcare applications. This chapter focuses on different probiotic bacteria and their beneficial effects, derivatives of probiotic bacteria responsible for those effects, different methodologies of synthesizing PBC by combining BC and probiotic bacteria and their derivatives, and the applications of PBC in different biomedical applications.